Solid forms of n-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide

ABSTRACT

The present invention relates to solid state forms of N-[2,4-bis(1,1-diethylethyl)-5-hydroxyphenyl]-1,4-dihydro-1-oxoquinoline-3-carboxamide (Compound 1), pharmaceutical compositions thereof and methods therewith.

CLAIM OF PRIORITY

This application claims priority under 35 USC § 119(e) to U.S. PatentApplication Ser. No. 60/754,381, filed on Dec. 28, 2005, the entirecontents of which are hereby incorporated by reference.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to solid state forms, for example,crystalline and amorphous forms, ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide,pharmaceutical compositions thereof, and methods therewith.

BACKGROUND OF THE INVENTION

CFTR is a cAMP/ATP-mediated anion channel that is expressed in a varietyof cells types, including absorptive and secretory epithelia cells,where it regulates anion flux across the membrane, as well as theactivity of other ion channels and proteins. In epithelia cells, normalfunctioning of CFTR is critical for the maintenance of electrolytetransport throughout the body, including respiratory and digestivetissue. CFTR is composed of approximately 1480 amino acids that encode aprotein made up of a tandem repeat of transmembrane domains, eachcontaining six transmembrane helices and a nucleotide binding domain.The two transmembrane domains are linked by a large, polar, regulatory(R)-domain with multiple phosphorylation sites that regulate channelactivity and cellular trafficking.

The gene encoding CFTR has been identified and sequenced (See Gregory,R. J. et al. (1990) Nature 347:382-386; Rich, D. P. et al. (1990) Nature347:358-362), (Riordan, J. R. et al. (1989) Science 245:1066-1073). Adefect in this gene causes mutations in CFTR resulting in cysticfibrosis (“CF”), the most common fatal genetic disease in humans. Cysticfibrosis affects approximately one in every 2,500 infants in the UnitedStates. Within the general United States population, up to 10 millionpeople carry a single copy of the defective gene without apparent illeffects. In contrast, individuals with two copies of the CF associatedgene suffer from the debilitating and fatal effects of CF, includingchronic lung disease.

In patients with cystic fibrosis, mutations in CFTR endogenouslyexpressed in respiratory epithelia leads to reduced apical anionsecretion causing an imbalance in ion and fluid transport. The resultingdecrease in anion transport contributes to enhanced mucus accumulationin the lung and the accompanying microbial infections that ultimatelycause death in CF patients. In addition to respiratory disease, CFpatients typically suffer from gastrointestinal problems and pancreaticinsufficiency that, if left untreated, results in death. In addition,the majority of males with cystic fibrosis are infertile and fertilityis decreased among females with cystic fibrosis. In contrast to thesevere effects of two copies of the CF associated gene, individuals witha single copy of the CF associated gene exhibit increased resistance tocholera and to dehydration resulting from diarrhea—perhaps explainingthe relatively high frequency of the CF gene within the population.

Sequence analysis of the CFTR gene of CF chromosomes has revealed avariety of disease causing mutations (Cutting, G. R. et al. (1990)Nature 346:366-369; Dean, M. et al. (1990) Cell 61:863:870; and Kerem,B-S. et al. (1989) Science 245:1073-1080; Kerem, B-S et al. (1990) Proc.Natl. Acad. Sci. USA 87:8447-8451). To date, >1000 disease causingmutations in the CF gene have been identified(http://www.genet.sickkids.on.ca/cftr/). The most prevalent mutation isa deletion of phenylalanine at position 508 of the CFTR amino acidsequence, and is commonly referred to as ΔF508-CFTR. This mutationoccurs in approximately 70% of the cases of cystic fibrosis and isassociated with a severe disease.

The deletion of residue 508 in ΔF508-CFTR prevents the nascent proteinfrom folding correctly. This results in the inability of the mutantprotein to exit the ER, and traffic to the plasma membrane. As a result,the number of channels present in the membrane is far less than observedin cells expressing wild-type CFTR. In addition to impaired trafficking,the mutation results in defective channel gating. Together, the reducednumber of channels in the membrane and the defective gating lead toreduced anion transport across epithelia leading to defective ion andfluid transport. (Quinton, P. M. (1990), FASEB J. 4: 2709-2727). Studieshave shown, however, that the reduced numbers of ΔF508-CFTR in themembrane are functional, albeit less than wild-type CFTR. (Dalemans etal. (1991), Nature Lond. 354: 526-528; Denning et al., supra; Pasyk andFoskett (1995), J. Cell. Biochem. 270: 12347-50). In addition toΔF508-CFTR, other disease causing mutations in CFTR that result indefective trafficking, synthesis, and/or channel gating could be up- ordown-regulated to alter anion secretion and modify disease progressionand/or severity.

Although CFTR transports a variety of molecules in addition to anions,it is clear that this role (the transport of anions) represents oneelement in an important mechanism of transporting ions and water acrossthe epithelium. The other elements include the epithelial Na⁺ channel,ENaC, Na⁺/2Cl⁻/K⁺ co-transporter, Na⁺—K⁺-ATPase pump and the basolateralmembrane K⁺ channels, that are responsible for the uptake of chlorideinto the cell.

These elements work together to achieve directional transport across theepithelium via their selective expression and localization within thecell. Chloride absorption takes place by the coordinated activity ofENaC and CFTR present on the apical membrane and the Na⁺—K⁺-ATPase pumpand Cl— channels expressed on the basolateral surface of the cell.Secondary active transport of chloride from the luminal side leads tothe accumulation of intracellular chloride, which can then passivelyleave the cell via Cl⁻ channels, resulting in a vectorial transport.Arrangement of Na⁺/2Cl⁻/K⁺ co-transporter, Na⁺—K⁺-ATPase pump and thebasolateral membrane K channels on the basolateral surface and CFTR onthe lunminal side coordinate the secretion of chloride via CFTR on theluminal side. Because water is probably never actively transporteditself, its flow across epithelia depends on tiny transepithelialosmotic gradients generated by the bulk flow of sodium and chloride.

In addition to cystic fibrosis, modulation of CFTR activity may bebeneficial for other diseases not directly caused by mutations in CFTR,such as secretory diseases and other protein folding diseases mediatedby CFTR. These include, but are not limited to, chronic obstructivepulmonary disease (COPD), dry eye disease, and Sjögren's Syndrome. COPDis characterized by airflow limitation that is progressive and not fullyreversible. The airflow limitation is due to mucus hypersecretion,emphysema, and bronchiolitis. Activators of mutant or wild-type CFTRoffer a potential treatment of mucus hypersecretion and impairedmucociliary clearance that is common in COPD. Specifically, increasinganion secretion across CFTR may facilitate fluid transport into theairway surface liquid to hydrate the mucus and optimized periciliaryfluid viscosity. This would lead to enhanced mucociliary clearance and areduction in the symptoms associated with COPD. Dry eye disease ischaracterized by a decrease in tear aqueous production and abnormal tearfilm lipid, protein and mucin profiles. There are many causes of dryeye, some of which include age, Lasik eye surgery, arthritis,medications, chemical/thermal burns, allergies, and diseases, such ascystic fibrosis and Sjögrens's syndrome. Increasing anion secretion viaCFTR would enhance fluid transport from the corneal endothelial cellsand secretory glands surrounding the eye to increase corneal hydration.This would help to alleviate the symptoms associated with dry eyedisease. Sjögrens's syndrome is an autoimmune disease in which theimmune system attacks moisture-producing glands throughout the body,including the eye, mouth, skin, respiratory tissue, liver, vagina, andgut. Symptoms, include, dry eye, mouth, and vagina, as well as lungdisease. The disease is also associated with rheumatoid arthritis,systemic lupus, systemic sclerosis, and polymypositis/dermatomyositis.Defective protein trafficking is believed to cause the disease, forwhich treatment options are limited. Modulators of CFTR activity mayhydrate the various organs afflicted by the disease and help to elevatethe associated symptoms.

As discussed above, it is believed that the deletion of residue 508 inΔF508-CFTR prevents the nascent protein from folding correctly,resulting in the inability of this mutant protein to exit the ER, andtraffic to the plasma membrane. As a result, insufficient amounts of themature protein are present at the plasma membrane and chloride transportwithin epithelial tissues is significantly reduced. Infact, thiscellular phenomenon of defective ER processing of ABC transporters bythe ER machinery, has been shown to be the underlying basis not only forCF disease, but for a wide range of other isolated and inheriteddiseases. The two ways that the ER machinery can malfunction is eitherby loss of coupling to ER export of the proteins leading to degradation,or by the ER accumulation of these defective/misfolded proteins [AridorM, et al., Nature Med., 5(7), pp 745-751 (1999); Shastry, B. S., et al.,Neurochem. International, 43, pp 1-7 (2003); Rutishauser, J., et al.,Swiss Med Wkly, 13, pp 211-222 (2002); Morello, J P et al., TIPS, 21 pp.466-469 (2000); Bross P., et al., Human Mut., 1, pp. 186-198 (1999)].The diseases associated with the first class of ER malfunction arecystic fibrosis (due to misfolded ΔF508-CFTR as discussed above),hereditary emphysema (due to al-antitrypsin; non Piz variants),hereditary hemochromatosis, hoagulation-fibrinolysis deficiencies, suchas protein C deficiency, Type 1 hereditary angioedema, lipid processingdeficiencies, such as familial hypercholesterolemia, Type 1chylomicronemia, abetalipoproteinemia, lysosomal storage diseases, suchas I-cell disease/pseudo-Hurler, Mucopolysaccharidoses (due to lysosomalprocessing enzymes), Sandhof/Tay-Sachs (due to β-hexosaminidase),Crigler-Najjar type II (due to UDP-glucuronyl-sialyc-transferase),polyendocrinopathy/hyperinsulemia, Diabetes mellitus (due to insulinreceptor), Laron dwarfism (due to growth hormone receptor),myleoperoxidase deficiency, primary hypoparathyroidism (due topreproparathyroid hormone), melanoma (due to tyrosinase). The diseasesassociated with the latter class of ER malfunction are Glycanosis CDGtype 1, hereditary emphysema (due to al-Antitrypsin (PiZ variant),congenital hyperthyroidism, osteogenesis imperfecta (due to Type I, II,IV procollagen), hereditary hypofibrinogenemia (due to fibrinogen), ACTdeficiency (due to α1-antichymotrypsin), Diabetes insipidus (DI),neurophyseal DI (due to vasopvessin hormone/V2-receptor), neprogenic DI(due to aquaporin II), Charcot-Marie Tooth syndrome (due to peripheralmyelin protein 22), Perlizaeus-Merzbacher disease, neurodegenerativediseases such as Alzheimer's disease (due to βAPP and presenilins),Parkinson's disease, amyotrophic lateral sclerosis, progressivesupranuclear plasy, Pick's disease, several polyglutamine neurologicaldisorders a such as Huntington, spinocerebullar ataxia type I, spinaland bulbar muscular atrophy, dentatorubal pallidoluysian, and myotonicdystrophy, as well as spongiform encephalopathies, such as hereditaryCreutzfeldt-Jakob disease (due to prion protein processing defect),Fabry disease (due to lysosomal α-galactosidase A) andStraussler-Scheinker syndrome (due to Prp processing defect).

In addition to up-regulation of CFTR activity, reducing anion secretionby CFTR modulators may be beneficial for the treatment of secretorydiarrheas, in which epithelial water transport is dramatically increasedas a result of secretagogue activated chloride transport. The mechanisminvolves elevation of cAMP and stimulation of CFTR.

Although there are numerous causes of diarrhea, the major consequencesof diarrheal diseases, resulting from excessive chloride transport arecommon to all, and include dehydration, acidosis, impaired growth anddeath.

Acute and chronic diarrheas represent a major medical problem in manyareas of the world. Diarrhea is both a significant factor inmalnutrition and the leading cause of death (5,000,000 deaths/year) inchildren less than five years old.

Secretory diarrheas are also a dangerous condition in patients ofacquired immunodeficiency syndrome (AIDS) and chronic inflammatory boweldisease (IBD). 16 million travelers to developing countries fromindustrialized nations every year develop diarrhea, with the severityand number of cases of diarrhea varying depending on the country andarea of travel.

Diarrhea in barn animals and pets such as cows, pigs and horses, sheep,goats, cats and dogs, also known as scours, is a major cause of death inthese animals. Diarrhea can result from any major transition, such asweaning or physical movement, as well as in response to a variety ofbacterial or viral infections and generally occurs within the first fewhours of the animal's life.

The most common diarrheal causing bacteria is enterotoxogenic E. coli(ETEC) having the K99 pilus antigen. Common viral causes of diarrheainclude rotavirus and coronavirus. Other infectious agents includecryptosporidium, giardia lamblia, and salmonella, among others.

Symptoms of rotaviral infection include excretion of watery feces,dehydration and weakness. Coronavirus causes a more severe illness inthe newborn animals, and has a higher mortality rate than rotaviralinfection. Often, however, a young animal may be infected with more thanone virus or with a combination of viral and bacterial microorganisms atone time. This dramatically increases the severity of the disease.

Accordingly, there is a need for stable polymorphic forms of modulatorsof CFTR activity, such as Compound 1, that can be used to modulate theactivity of CFTR in the cell membrane of a mammal.

There is a need for methods of treating CFTR-mediated diseases usingsuch modulators of CFTR activity.

SUMMARY OF THE INVENTION

The present invention relates to solid forms ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide(hereinafter “Compound 1”) which has the structure below:

The solid forms of Compound 1 and pharmaceutically acceptablecompositions thereof are useful for treating or lessening the severityof a variety of CFTR mediated diseases. Compound 1 is known as bothN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideandN-(5-hydroxy-2,4-di-tert-butyl-phenyl)-4-oxo-1H-quinoline-3-carboxamide.

In one aspect, the invention features solid amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.In some embodiments, the solid amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidecomprises less than about 15% crystallineN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

In one aspect, the invention features a preparation of amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidesubstantially free of crystallineN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

In some embodiments, the preparation further comprises a surfactant,polymer, or inert pharmaceutically acceptable substance.

In some embodiments, the preparation comprises a solid dispersion, amixture or a liquid dispersion.

In some embodiments, the preparation comprises solid particles.

In some embodiments, the preparation comprises less than about 15% ofcrystallineN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

In some embodiments, the amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidehas a particle size distribution of D10, less than 5 μm. In someembodiments, the amporphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidehave a particle size distribution of D50, less than 17 μm. In someembodiments, the amporphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidehave a particle size distribution of D90, less than 100 μm.

In one aspect, the invention features a solid dispersion comprisingamorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

In some embodiments, the solid dispersion comprises less than about 40%of crystallineN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.In some embodiments, the solid dispersion is substantially free ofcrystallineN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

In some embodiments, the solid dispersion further comprises asurfactant, polymer, or inert pharmaceutically acceptable substance. Forexample, the solid dispersion comprises a polymer, and the polymer isone or more than one water-soluble polymer or partially water-solublepolymer.

In some embodiments, theN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidehas improved physical or chemical stability relative to amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidewithout being in the presence of polymer.

In some embodiments, the solid dispersion has a higher glass transitiontemperature than the glass transition temperature of neat amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

In some embodiments, the polymer is hydroxypropylmethylcellulose (HPMC).In some embodiments, the polymer is hydroxypropylmethylcellulose acetatesuccinate (HPMCAS).

In some embodiments, the polymer is vinylpyrrolidone/vinyl acetatecopolymer (PVP/VA). In some embodiments, the polymer is present in anamount of from about 10% by weight to about 80% by weight, for example,the polymer is present in an amount of less than about 70% by weight,the polymer is present in an amount of about 50% by weight, or thepolymer is present in an amount of about 49.5% by weight.

In some embodiments, theN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis present in an amount of from about 10% by weight to about 80% byweight, for example, theN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis present in an amount of less than about 70% by weight or theN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis present in an amount of about 50% by weight.

In some embodiments, the solid dispersion comprises a surfactant, forexample, sodium lauryl sulfate. In some embodiments, the surfactant ispresent in an amount from about 0.1 to about 5%, for example, thesurfactant is present in 0.5%.

In some embodiments, at least about 80% by weight of the NN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis in an amorphous form. In some embodiments, substantially all theN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis in an amorphous form.

In some embodiments, the solid dispersion is obtained by spray drying.

In one aspect, the invention features a pharmaceutical compositioncomprising amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.In some embodiments, the amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis substantially free of crystallineN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

In one aspect, the invention features, a pharmaceutical compositioncomprising an amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideas a solid dispersion and one or more of a surfactant, polymer, inertpharmaceutically acceptable substance, or pharmaceutically acceptablecarrier.

In some embodiments, the solid dispersion comprises a polymer andwherein the polymer is one or more than one water-soluble polymer orpartially water-soluble polymer.

In some embodiments, the solid dispersion has a higher glass transitiontemperature than the glass transition temperature of neat amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

In some embodiments, the polymer is HPMC. In some embodiments, thepolymer is HPMCAS. In some embodiments, the polymer is PVP/VA.

In one aspect, the invention features a pharmaceutical compositioncomprising:

an amorphous solid dispersion ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidewherein saidN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidecomprises about 30-75% wt/wt of the pharmaceutical composition, one ormore polymer selected from the group of HPMC and HPMCAS, wherein saidpolymer is comprises about 30-75% wt/wt of the pharmaceuticalcomposition, and a surfactant, wherein said surfactant comprises about0.25-1% wt/wt of the pharmaceutical composition.

In some embodiments, the polymer is HPMCAS. In some embodiments, thepolymer is HPMC.

In some embodiments, the surfactant is sodium laurel sulfate.

In some embodiments, saidN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidecomprises about 50% wt/wt of the pharmaceutical composition, saidpolymer is HPMCAS and comprises about 49.5% wt/wt of the pharmaceuticalcomposition, and a said surfactant is sodium laurel sulfate andcomprises about 0.5% wt/wt of the pharmaceutical composition.

In one aspect, the invention features a pharmaceutical compositioncomprising;

an aqueous suspension comprising amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideparticles and a pharmaceutically acceptable carrier.

In some embodiments, the pharmaceutically acceptable carrier is apolymer in solution selected from the group of HPMC and HPMCAS. In someembodiments, the pharmaceutically acceptable carrier is a polymer insolution is PVP/VA.

In some embodiments, the amorphous compound is in the form of a soliddispersion.

In some embodiments, the pharmaceutical composition further comprises asurfactant, either in the solution or as a component of the soliddispersion, for example, SLS. In some embodiments, the polymer is eitherin the solution or as a component of the solid dispersion particles orboth. In some embodiments, the aqueous suspension comprises from about0.1% to about 20% by weight of the surfactant. In some embodiments, theaqueous suspension comprises from about 0.1% to about 2.0% by weight ofpolymer, for example, about 1% by weight of polymer.

In one aspect, the invention features a process for preparing anamorphous form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidecomprising spray-dryingN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideto provide an amorphous form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

In some embodiments, the method comprises combiningN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideand a suitable solvent to form a mixture and then spray-drying themixture to obtain the amorphous form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl)-1,4-dihydro-4-oxoquinoline-3-carboxamide.

In some embodiments, the mixture is a solutionN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideand the suitable solvent. In some embodiments, the suitable solventcomprises acetone or MEK. In some embodiments, the suitable solventcomprises a mixture of solvents, for example, a mixture of acetone andwater or a mixture of MEK and water. In some embodiments, the water inthe solvent mixture is present at about 10% wt.

In some embodiments, the method comprises a) forming a mixturecomprisingN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide,a polymer, and a solvent; and b) spray-drying the mixture to form asolid dispersion comprisingN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

In some embodiments, the mixture comprises a solution ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide,the polymer, and the solvent. In some embodiments, the polymer isselected from HPMC and HPMCAS. In some embodiments, the polymer isPVP/VA. In some embodiments, the polymer is present in an amount of fromabout 30% to about 70% by weight in the solid dispersion. In someembodiments, the mixture further comprises a surfactant, for example,SLS.

In some embodiments, the solvent comprises acetone, for example, amixture of acetone and water. In some embodiments, the solvent comprisesfrom about 0% to about 20% water and from about 70% to about 100%acetone.

In one aspect, the invention features a solid dispersion preparedaccording a process described herein.

In one aspect, the invention features a method for treating aCFTR-mediated disease in a mammal comprising administering amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.In some embodiments, the method comprises administering an amorphoussolid dispersion ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.In some embodiments, the method comprises administering an additionaltherapeutic agent.

In one aspect, the invention features a pharmaceutical pack or kitcomprising amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideand a pharmaceutically acceptable carrier.

In one aspect, the invention features a crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide,characterized by one or more peaks at from about 4.8 to about 5.2degrees, for example, about 5.0 degrees, and from about 15.4 to about15.8 degrees, for example, about 15.6 degrees in an X-ray powderdiffraction pattern obtained using Cu K alpha radiation. In someembodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak at from about 7.6 toabout 8.0, e.g., 7.8. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak at from about 8.3 toabout 8.7, for example, about 8.5. In some embodiments, the crystal formofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak at from about 9.0 toabout 9.4, for example, about 9.2. In some embodiments, the crystal formofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak at from about 9.7 toabout 10.1, for example about 9.9. In some embodiments, the crystal formofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak at from about 11.7 toabout 12.1, for example, about 11.9. In some embodiments, the crystalform ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak at from about 12.4 toabout 12.8, for example, about 12.6. In some embodiments, the crystalform ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak at from about 13.7 toabout 14.1, for example about 13.9. In some embodiments, the crystalform ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak at from about 14.7 toabout 15.1, for example, about 14.9. In some embodiments, the crystalform ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 16.3 to about16.7, for example about 16.5. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 17.9 to about18.3, for example, about 18.1. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 18.3 to about18.7, for example, about 18.5. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-diydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 20.5 to about20.9, for example, about 20.7. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 21.8 to about22.2, for example, about 22.0. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 23.1 to about23.7, for example, about 23.5. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 25.1 to about25.5, for example, about 25.3. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 27.8 to about28.2, for example, about 28.0. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 29.2 to about29.6, for example, about 29.4. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 30.7 to about31.1, for example, about 30.9. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis characterized by an X-ray powder diffraction pattern obtained usingCu K alpha radiation substantially similar to FIG. 4.

In one aspect, the invention features a pharmaceutical compositioncomprising the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidehaving the characteristics of Form A, for example as described above,and a pharmaceutically acceptable adjuvant or carrier.

In one aspect, the invention features a process for preparing a crystalform ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideof Form A, for example as as characterized above, wherein said processcomprises the step of heating NN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideto about 250° C. and cooling to room temperature.

In one aspect, the invention features a method for treating a CFTRmediated disease in a mammal comprising administeringN-[2,4-bis(1,1-dimethylethy)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideof Form A, for example as as characterized above.

In some embodiments, theN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis a component of a pharmaceutical composition. In some embodiments, themethod comprises administering an additional therapeutic agent.

In one aspect, the invention features a pharmaceutical pack or kitcomprising crystallineN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideof Form A, for example as as characterized above and a pharmaceuticallyacceptable carrier.

In one aspect, the invention features a crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide,characterized by one or more peaks at from about 6.2 to about 6.6, forexample, about 6.4, from about 7.5 to about 7.9, for example, about 7.7,from about 12.5 to about 12.9, for example, about 12.7, and from about17.9 to about 18.3, for example, about 18.1 degrees in an X-ray powderdiffraction pattern obtained using Cu K alpha radiation.

In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 8.2 to about8.6, for example, about 8.4. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 10.8 to about11.2, for example, about 11.0. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 14.6 to about15.0, for example, about 14.8. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 15.9 to about16.3, for example, about 16.1. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 16.9 to about17.3, for example, about 17.1. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 18.4 to about18.8, for example, about 18.6. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 19.2 to about19.6, for example, about 19.4. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 20.9 to about21.3, for example, about 21.1. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 22.4 to about22.8, for example, about 22.6. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 23.2 to about23.6, for example, about 23.4. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 23.7 to about24.1, for example, about 23.9. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 24.7 to about25.1, for example, about 24.9. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 25.3 to about25.7, for example, about 25.5. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 26.5 to about26.9, for example, about 26.7. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 27.3 to about27.7, for example, about 27.5. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis further characterized by the following peak from about 29.4 to about29.8, for example, about 29.6. In some embodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro4-oxoquinoline-3-carboxamide is further characterized by the followingpeak from about 33.3 to about 33.7, for example, about 33.5. In someembodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro4-oxoquinoline-3-carboxamide is further characterized by the followingpeak from about 36.6 to about 37.0, for example, about 36.8. In someembodiments, the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideis characterized by an X-ray powder diffraction pattern obtained usingCu K alpha radiation substantially similar to FIG. 7.

In one aspect, the invention features a crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide,having a monoclinic crystal system, a P21 space grouping, and thefollowing unit cell dimensions:

a=11.8011(7) Å α=90°

b=5.9819(3) Å β=105.110(4°)

c=14.7974(8) Å γ=90°.

In one aspect, the invention features a pharmaceutical compositioncomprising the crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideaccording to Form B, for example, as characterized above, and apharmaceutically acceptable adjuvant or carrier.

In one aspect, the invention features a process for preparing thecrystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquincline-3-carboxamideaccording to Form B, for example, as characterized above, wherein saidprocess comprises the steps of alternatively heating and cooling aslurry of Compound 1 and acetonitrile. In some embodiments, the processcomprises heating said slurry at about 50 C for about 12 hours. In someembodiments, said cooling step comprises placing said slurry at roomtemperature for about 12 hours, followed by cooling at about 0° C.overnight.

In one aspect, the invention features a method of treating a CFTRmediated disease in a patient comprising the step of administering tosaid patient a crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideor a pharmaceutical composition comprising a crystal form ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideaccording to Form B, for example, as characterized above.

In one aspect, the invention features a method of treating a disease isselected from cystic fibrosis, hereditary emphysema, hereditaryhemochromatosis, coagulation-fibrinolysis deficiencies, such as proteinC deficiency, Type 1 hereditary angioedema, lipid processingdeficiencies, such as familial hypercholesterolemia, Type 1chylomicronemia, abetalipoproteinemia, lysosomal storage diseases, suchas I-cell disease/pseudo-Hurler, mucopolysaccharidoses,Sandhof/Tay-Sachs, Crigler-Najjar type I,polyendocrinopathy/hyperinsulemia, Diabetes mellitus, Laron dwarfism,myleoperoxidase deficiency, primary hypoparathyroidism, melanoma,glycanosis CDG type 1, hereditary emphysema, congenital hyperthyroidism,osteogenesis imperfecta, hereditary hypofibrinogenemia, ACT deficiency,Diabetes insipidus (DI), neurophyseal DI, neprogenic DI, Charcot-MarieTooth syndrome, Perlizaeus-Merzbacher disease, neurodegenerativediseases such as Alzheimer's disease, Parkinson's disease, amyotrophiclateral sclerosis, progressive supranuclear plasy, Pick's disease,several polyglutamine neurological disorders asuch as Huntington,spinocerebullar ataxia type I, spinal and bulbar muscular atrophy,dentatorubal pallidoluysian, and myotonic dystrophy, as well asspongiform encephalopathies, such as hereditary Creutzfeldt-Jakobdisease, Fabry disease, Straussler-Scheinker syndrome, COPD, dry-eyedisease, and Sjogren's disease by administering a solid form of Compound1 as described above, for example, Form A, Form B, or amorphous Compound1, for example, neat or as a component of a solid dispersion. In someembodiments, the disease is cystic fibrosis.

Processes described herein can be used to prepare the compositions ofthis invention. The amounts and the features of the components used inthe processes would be as described herein.

As used herein, the term “amorphous” refers to a solid material havingno long range order in the position of its molecules Amorphous solidsare generally supercooled liquids in which the molecules are arranged ina random manner so that there is no well-defined arrangement, e.g.,molecular packing, and no long range order. Amorphous solids aregenerally isotropic, i.e. exhibit similar properties in all directionsand do not have definite melting points. For example, an amorphousmaterial is a solid material having no sharp characteristic crystallinepeak(s) in its X-ray power diffraction (XRPD) pattern (i.e., is notcrystalline as determined by XRPD). Instead, one or several broad peaks(e.g., halos) appear in its XRPD pattern. Broad peaks are characteristicof an amorphous solid. See, US 2004/0006237 for a comparison of XRPDs ofan amorphous material and crystalline material.

As used herein, the phrase “substantially amorphous Compound 1” is usedinterchangeably with the phrase “amorphous Compound 1 substantially freeof crystalline Compound 1.” In some embodiments, substantially amorphousCompound 1 has less than about 30% crystalline Compound 1, for example,less than about 30% of crystalline Compound 1, e.g., less than about 25%crystalline Compound 1, less than about 20% crystalline Compound 1, lessthan about 15% crystalline Compound 1, less than about 10% crystallineCompound 1, less than about 5% crystalline Compound 1, less than about2% crystalline Compound 1. In some preferred embodiments, Compound 1 hasless than about 15% crystalline compound 1. Some embodiments include apreparation of substantially amorphous Compound 1, for example havingthe degree of crystalline Compound 1 as described above.

As used herein “crystalline solids” refers to compounds or compositionswhere the structural units are arranged in fixed geometric patterns orlattices, so that crystalline solids have rigid long range order. Thestructural units that constitute the crystal structure can be atoms,molecules, or ions. Crystalline solids show definite melting points.

As used herein, a “dispersion” refers to a disperse system in which onesubstance, the dispersed phase, is distributed, in discrete units,throughout a second substance (the continuous phase or vehicle). Thesize of the dispersed phase can vary considerably (e.g. colloidalparticles of nanometer dimension, to multiple microns in size). Ingeneral, the dispersed phases can be solids, liquids, or gases. In thecase of a solid dispersion, the dispersed and continuous phases are bothsolids. In pharmaceutical applications, a solid dispersion can include acrystalline drug (dispersed phase) in an amorphous polymer (continuousphase), or alternatively, an amorphous drug (dispersed phase) in anamorphous polymer (continuous phase). In some embodiments an amorphoussolid dispersion includes the polymer constituting the dispersed phase,and the drug constitute the continuous phase. In some embodiments, thedispersion includes amorphous Compound 1 or substantially amorphousCompound 1.

The term “solid amorphous dispersion” generally refers to a soliddispersion of two or more components, usually a drug and polymer, butpossibly containing other components such as surfactants or otherpharmaceutical excipients, where Compound 1 is amorphous orsubstantially amorphous (e.g., substantially free of crystallineCompound 1), and the physical stability and/or dissolution and/orsolubility of the amorphous drug is enhanced by the other components.

A solid dispersion as provided herein is a particularly favorableembodiment of this invention. Solid dispersions typically include acompound dispersed in an appropriate carrier medium, such as a solidstate carrier. In one embodiment, a carrier according to this inventioncomprises a polymer, preferably, a water-soluble polymer or a partiallywater-soluble polymer. It would be understood that one or more than onewater-soluble polymer could be used in a solid dispersion of thisinvention.

An exemplary solid dispersion is a co-precipitate or a co-melt ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidewith at least one polymer. A “Co-precipitate” is a product afterdissolving a drug and a polymer in a solvent or solvent mixture followedby the removal of the solvent or solvent mixture. Sometimes the polymercan be suspended in the solvent or solvent mixture. The solvent orsolvent mixture includes organic solvents and supercritical fluids. A“co-melt” is a product after heating a drug and a polymer to melt,optionally in the presence of a solvent or solvent mixture, followed bymixing, removal of at least a portion of the solvent if applicable, andcooling to room temperature at a selected rate. In some cases, the soliddispersions are prepared by adding a solution of a drug and a solidpolymer followed by mixing and removal of the solvent. To remove thesolvent, vacuum drying, spray drying, tray drying, lyophilization, andother drying procedures may be applied. Applying any of these methodsusing appropriate processing parameters, according to this invention,would provideN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidein an amorphous state in the final solid dispersion product.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an X-Ray powder diffraction pattern of Compound 1.

FIG. 2 is the ¹H NMR spectrum of Compound 1.

FIG. 3 is the DSC trace of Compound 1.

FIG. 4 is the X-Ray powder diffraction pattern of Form A.

FIG. 5 is the DSC trace of Form A.

FIG. 6 is the TGA trace of Form A.

FIG. 7 is the X-Ray powder diffraction pattern of Form B.

FIG. 8 is the DSC trace of Form B.

FIG. 9 is the TGA trace of Form B.

FIG. 10 is a conformational picture of Form B, based on single crystalX-Ray analysis.

FIG. 11 is the X-Ray powder diffraction pattern of the Amorphous Form.

FIG. 12 is the TGA trace of the Amorphous Form.

FIG. 13 is the DSC trace of the Amorphous Form.

DETAILED DESCRIPTION OF THE INVENTION

Solid Forms of Compound 1

Form A

Form A of Compound 1 is characterized by one or more peaks at from about4.8 to about 5.2, for example, about 5.0, e.g., 4.99, and from about15.4 to about 15.8, for example, about 15.6 e.g., 15.58 degrees in anX-ray powder diffraction pattern obtained using Cu K alpha radiation(20). Other peaks (20), which can be characteristic of Form A, includethe following: from about 7.6 to about 8.0, for example, about 7.8,e.g., 7.75; from about 8.3 to about 8.7, for example, about 8.5, e.g.,8.46; from about 9.0 to about 9.4, for example, about 9.2, e.g., 9.21;from about 9.7 to about 10.1, for example, about 9.9, e.g., 9.92; fromabout 11.7 to about 12.1, for example, about 11.9, e.g., 11.93; fromabout 12.4 to about 12.8, for example, about 12.6, e.g., 12.64; fromabout 13.7 to about 14.1, for example, about 13.9, e.g., 13.88; fromabout 14.7 to about 15.1, for example, about 14.9, e.g., 14.91; fromabout 16.3 to about 16.7, for example, about 16.5, e.g., 16.46; fromabout 17.9 to about 18.3, for example, about 18.1, e.g., 18.09; fromabout 18.3 to about 18.7, for example, about 18.5, e.g., 18.52; fromabout 21.5 to about 21.9, for example, about 21.7, e.g., 20.65; fromabout 21.8 to about 22.2, for example, about 22.0, e.g., 21.95; fromabout 23.1 to about 23.7, for example, about 23.5, e.g., 23.49; fromabout 25.1 to about 25.5, for example, about 25.3, e.g., 25.26; fromabout 27.8 to about 28.2 for example, about 28.0, e.g., 28.02; fromabout 29.2 to about 29.6, for example, about 29.4, e.g., 29.35; andabout from about 30.7 to about 31.1, for example, 30.9, e.g., 30.85. Forexample, Form A can be characterized by an X-ray powder diffractionpattern obtained using Cu K alpha radiation substantially similar toFIG. 4.

Pharmaceutical compositions including Form A and a pharmaceuticallyacceptable adjuvant or carrier, such as a polymer or surfactant are alsodescribed. Form A can be formulated in a pharmaceutical composition, insome instances, with another therapeutic agent, for example anothertherapeutic agent for treating cystic fibrosis or a symptom thereof.

Processes for preparing Form A are exemplified herein.

Methods of treating a CFTR mediated disease, such as cystic fibrosis, ina patient include administering to said patient Form A or apharmaceutical composition comprising Form A.

Form B

The solid state crystal Form B of Compound 1 is characterized by one ormore peaks at from about 6.0 to about 6.4 for example, about 6.2, e.g.,6.17, from about 7.4 to about 7.8 for example, about 7.6, e.g., 7.61,from about 12.1 to about 12.5 for example, about 12.3, e.g., 12.33, andfrom about 17.8 to about 18.2 for example, about 18.0, e.g., 17.96degrees in an X-ray powder diffraction pattern obtained using Cu K alpharadiation (20). Other peaks (20), which can be characteristic of Form B,include the following: from about 8.2 to about 8.6 for example, about8.4, e.g., 8.40; from about 10.8 to about 11.2 for example, about 11.0,e.g., 11.02; from about 14.6 to about 15.0 for example, about 14.8,e.g., 14.83; from about 15.9 to about 16.3 for example, about 16.1,e.g., 16.14; from about 16.9 to about 17.3 for example, about 17.1,e.g., 17.11; from about 18.4 to about 18.8 for example, about 18.6,e.g., 18.55; from about 19.2 to about 19.6 for example, about 19.4,e.g., 19.43; from about 20.9 to about 21.3 for example, about 21.1,e.g., 21.05; from about 22.4 to about 22.8 for example, about 22.6,e.g., 22.56; from about 23.2 to about 23.6 for example, about 23.4,e.g., 23.37; from about 23.7 to about 24.1 for example, about 23.9,e.g., 23.94; from about 24.7 to about 25.1 for example, about 24.9,e.g., 24.86; from about 25.3 to about 25.7 for example, about 25.5,e.g., 25.50; from about 26.5 to about 26.9 for example, about 26.7,e.g., 26.72; from about 27.3 to about 27.7 for example, about 27.5,e.g., 27.51; from about 29.4 to about 29.8 for example, about 29.6,e.g., 29.60; from about 33.3 to about 33.7 for example, about 33.5,e.g., 33.48; and from about 36.6 to about 37.0 for example, about 36.8,e.g., 36.78. Form B can be further characterized, for example, by anX-ray powder diffraction pattern obtained using Cu K alpha radiationsubstantially similar to FIG. 7.

Applicants have determined crystal structure dimensions of Form B byanalysis of single crystal data. Form B is a monoclinic crystal systemhaving a P2₁ space grouping, and the following unit cell dimensions:a=11.8011(7) Å, α=90°; b=5.9819(3) Å, β=105.110(4)° 1; c=14.7974(8) Å,γ=90°. Additional details about the structure and packing of Form B areprovided in the Examples.

Pharmaceutical compositions including Form B and a pharmaceuticallyacceptable adjuvant or carrier, such as a polymer or surfactant are alsodescribed. Form B can be formulated in a pharmaceutical composition, insome instances, with another therapeutic agent, for example anothertherapeutic agent for treating cystic fibrosis or a symptom thereof.

Processes for preparing Form B are exemplified herein.

Methods of treating a CFTR mediated disease, such as cystic fibrosis, ina patient include administering to said patient Form B or apharmaceutical composition comprising Form B.

Amorphous Compound 1

Compound 1 can be present as an amorpous solid, for example amorphousCompound 1 as a substantially neat preparation, or amorphous compound 1as a component as a dispersion such as a solid amorphous dispersion.

In some embodiments, an amorphous form of Compound 1 is substantiallyfree of crystalline Compound 1 (e.g., Form A, Form B or any crystallineform of Compound 1), for example Compound I has less than about 30% ofcrystalline Compound 1, e.g., less than about 25% crystalline Compound1, less than about 20% crystalline Compound 1, less than about 15%crystalline Compound 1, less than about 10% crystalline Compound 1, lessthan about 5% crystalline Compound 1, less than about 2% crystalline.Compound 1, preferably less than about 15% crystalline compound 1.Compound 1 can be characterized by an X-ray powder diffraction patternobtained using Cu K alpha radiation substantially similar to FIG. 11.For example, the substantially amorphous form of Compound 1 can becharacterized as having an XRPD having no sharp characteristiccrystalline peak(s) in its X-ray power diffraction (XRPD) pattern (i.e.,is not crystalline as determined by XRPD). Instead, one or several broadpeaks (e.g., halos) appear in its XRPD pattern.

Polymers

Solid dispersions including amorphous Compound 1 and a polymer (or solidstate carrier) also are included herein. For example, Compound 1 ispresent as an amorphous compound as a component of a solid amorphousdispersion. The solid amorphous dispersion, generally includes Compound1 and a polymer. Exemplary polymers include cellulosic polymers such asHPMC or HPMCAS and pyrrolidone containing polymers such as PVP/VA. Insome embodiments, the solid amporphous dispersion includes one or moreadditional excipients, such as a surfactant.

In one embodiment, a polymer is able to dissolve in aqueous media. Thesolubility of the polymers may be pH-independent or pH-dependent. Thelatter include one or more enteric polymers. The term “enteric polymer”refers to a polymer that is preferentially soluble in the less acidicenvironment of the intestine relative to the more acid environment ofthe stomach, for example, a polymer that is insoluble in acidic aqueousmedia but soluble when the pH is above 5-6. An appropriate polymershould be chemically and biologically inert. In order to improve thephysical stability of the solid dispersions, the glass transitiontemperature (T_(g)) of the polymer should be as high as possible. Forexample, preferred polymers have a glass transition temperature at leastequal to or greater than the glass transition temperature of the drug(e.g., Compound 1). Other preferred polymers have a glass transitiontemperature that is within about 10 to about 15° C. of the drug (e.g.,Compound 1). Examples of suitable glass transition temperatures of thepolymers include at least about 90° C., at least about 95° C., at leastabout 100° C., at least about 105° C., at least about 110° C., at leastabout 115° C., at least about 120° C., at least about 125° C., at leastabout 130° C., at least about 135° C., at least about 140° C., at leastabout 145° C., at least about 150° C., at least about 155° C., at leastabout 160° C., at least about 165° C., at least about 170° C., or atleast about 175° C. (as measured under dry conditions). Without wishingto be bound by theory, it is believed that the underlying mechanism isthat a polymer with a higher T_(g) generally has lower molecularmobility at room temperature, which can be a crucial factor instabilizing the physical stability of the amorphous solid dispersion.

Additionally, the hygroscopicity of the polymers should be as low, e.g.,less than about 10%. For the purpose of comparison in this application,the hygroscopicity of a polymer or composition is characterized at about60% relative humidity. In some preferred embodiments, the polymer hasless than about 10% water absorption, for example less than about 9%,less than about 8%, less than about 7%, less than about 6%, less thanabout 5%, less than about 4%, less than about 3%, or less than about 2%water absorption. The hygroscopicity can also affect the physicalstability of the solid dispersions. Generally, moisture adsorbed in thepolymers can greatly reduce the Ts of the polymers as well as theresulting solid dispersions, which will further reduce the physicalstability of the solid dispersions as described above.

In one embodiment, the polymer is one or more water-soluble polymer(s)or partially water-soluble polymer(s). Water-soluble or partiallywater-soluble polymers include but are not limited to, cellulosederivatives (e.g., hydroxypropylmethylcellulose (HPMC),hydroxypropylcellulose (HPC)) or ethylcellulose; polyvinylpyrrolidones(PVP); polyethylene glycols (PEG); polyvinyl alcohols (PVA); acrylates,such as polymethacrylate (e.g., Eudragit® E); cyclodextrins (e.g.,β-cyclodextin) and copolymers and derivatives thereof, including forexample PVP-VA (polyvinylpyrollidone-vinyl acetate).

In some preferred embodiments, the polymer ishydroxypropylmethylcellulose (HPMC), such as HPMC E50, HPMCE15, orHPMC60SH50).

As discussed herein, the polymer can be a pH-dependent enteric polymer.Such pH-dependent enteric polymers include, but are not limited to,cellulose derivatives (e.g., cellulose acetate phthalate (CAP)),hydroxypropyl methyl cellulose phthalates (HPMCP), hydroxypropyl methylcellulose acetate succinate (HPMCAS), carboxymethylcellulose (CMC) or asalt thereof (e.g., a sodium salt such as (CMC-Na)); cellulose acetatetrimellitate (CAT), hydroxypropylcellulose acetate phthalate (HPCAP),hydroxypropylmethyl-cellulose acetate phthalate (HPMCAP), andmethylcellulose acetate phthalate (MCAP), or polymethacrylates (e.g.,Eudragit® S). In some preferred embodiments, the polymer ishydroxypropyl methyl cellulose acetate succinate (HPMCAS).

In yet another embodiment, the polymer is a polyvinylpyrrolidoneco-polymer, for example, avinylpyrrolidone/vinyl acetate co-polymer(PVP/VA).

In embodiments where Compound 1 forms a solid dispersion with a polymer,for example with an HPMC, HPMCAS or PVP/VA polymer, the amount ofpolymer relative to the total weight of the solid dispersion ranges fromabout 0.1% to 99% by weight. Unless otherwise specified, percentages ofdrug, polymer and other excipients as described within a dispersion aregiven in weight percentages. The amount of polymer is typically at leastabout 20%, and preferably at least about 30%, for example, at leastabout 35%, at least about 40%, at least about 45%, or about 50% (e.g.,49.5%). The amount is typically about 99% or less, and preferably about80% or less, for example about 75% or less, about 70% or less, about 65%or less, about 60% or less, or about 55% or less. In one embodiment, thepolymer is in an amount of up to about 50% of the total weight of thedispersion (and even more specifically, between about 40% and 50%, suchas about 49%, about 49.5%, or about 50%). HPMC and HPMCAS are availablein a variety of grades from ShinEtsu, for example, HPMCAS is availablein a number of varieties, including AS-LF, AS-MF, AS-HF, AS-LG, AS-MG,AS-HG. Each of these grades vary with the percent substitution ofacetate and succinate.

In some preferred embodiments, Compound 1 and polymer are present inroughly equal amounts, for example each of the polymer and the drug makeup about half of the percentage weight of the dispersion. For example,the polymer is present in about 49.5% and the drug is present in about50%.

In some preferred embodiments, the dispersion further includes otherminor ingredients, such as a surfactant (e.g., SLS). In some preferredembodiments, the surfactant is present in less than about 10% of thedispersion, for example less than about 9%, less than about 8%, lessthan about 7%, less than about 6%, less than about 5%, less than about4%, less than about 3%, less than about 2%, about 1%, or about 0.5%.

In embodiments including a polymer, the polymer should be present in anamount effective for stabilizing the solid dispersion. Stabilizingincludes inhibiting or preventing, the crystallization of Compound 1.Such stabilizing would inhibit the conversion Compound 1 from amorphousto crystalline form. For example, the polymer would prevent at least aportion (e.g., about 5%, about 10%, about 15%, about 20%, about 25%,about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about60%, about 65%, about 70%, about 75%, or greater) of Compound 1 fromconverting from an amorphous to a crystalline form. Stabilization can bemeasured, for example, by measuring the glass transition temperature ofthe solid dispersion, measuring the rate of relaxation of the amorphousmaterial, or by measuring the solubility or bioavailability of Compound1.

Suitable polymers for use in combination with Compound 1, for example toform a solid dispersion such as an amorphous solid dispersion, shouldhave one or more of the following properties:

The glass transition temperature of the polymer should have atemperature of no less than about 10-15° C. lower than the glasstransition temperature of Compound 1. Preferably, the glass transitiontemperature of the polymer is greater than the glass transitiontemperature of Compound 1, and in general at least 50° C. higher thanthe desired storage temperature of the drug product. For example, atleast about 100° C., at least about 105° C., at least about 105° C., atleast about 110° C., at least about 120° C., at least about 130° C., atleast about 140° C., at least about 150° C., at least about 160° C., atleast about 160° C., or greater.

The polymer should be relatively non-hygroscopic. For example, thepolymer should, when stored under standard conditions, absorb less thanabout 10% water, for example, less than about 9%, less than about 8%,less than about 7%, less than about 6%, or less than about 5%, less thanabout 4%, or less than about 3% water. Preferably the polymer will, whenstored under standard conditions, be substantially free of absorbedwater.

The polymer should have similar or better solubility in solventssuitable for spray drying processes relative to that of Compound 1. Inpreferred embodiments, the polymer will dissolve in one or more of thesame solvents or solvent systems as Compound 1. It is preferred that thepolymer is soluble in at least one non-hydroxy containing solvent suchas methylene chloride, acetone, or a combination thereof.

The polymer, when combined with Compound 1, for example in a soliddispersion or in a liquid suspension, should increase the solubility ofCompound 1 in aqueous and physiologically relative media either relativeto the solubility of Compound 1 in the absence of polymer or relative tothe solubility of Compound 1 when combined with a reference polymer.

For example, the polymer could increase the solubility of amorphousCompound 1 by reducing the amount of amorphous Compound 1 that convertsto crystalline Compound 1, either from a solid amorphous dispersion orfrom a liquid suspension.

The polymer should decrease the relaxation rate of the amorphoussubstance.

The polymer should increase the physical and/or chemical stability ofCompound 1.

The polymer should improve the manufacturability of Compound 1.

The polymer should improve one or more of the handling, administrationor storage properties of Compound 1.

The polymer should not interact unfavorably with other pharmaceuticalcomponents, for example excipients.

The suitability of a candidate polymer (or other component) can betested using the spray drying methods (or other methods) describedherein to form an amorphous composition. The candidate composition canbe compared in terms of stability, resistance to the formation ofcrystals, or other properties, and compared to a reference preparation,e.g., a preparation of neat amorphous Compound 1 or crystallineCompound 1. E.g., a candidate composition could be tested to determinewhether it inhibits the time to onset of solvent mediatedcrystallization, or the percent conversion at a given time undercontrolled conditions, by at least 50%, 75%, 100%, or 110% as well asthe reference preparation, or a candidate composition could be tested todetermine if it has improved bioavailability or solubility relative tocrystalline Compound 1.

Surfactants

A solid dispersion or other composition may include a surfactant. Asurfactant or surfactant mixture would generally decrease theinterfacial tension between the solid dispersion and an aqueous medium.An appropriate surfactant or surfactant mixture may also enhance aqueoussolubility and bioavailability of Compound 1 from a solid dispersion.The surfactants for use in connection with the present inventioninclude, but are not limited to, sorbitan fatty acid esters (e.g.,Spans), polyoxyethylene sorbitan fatty acid esters (e.g., Tweens®),sodium lauryl sulfate (SLS), sodium dodecylbenzene sulfonate (SDBS)dioctyl sodium sulfosuccinate (Docusate), dioxycholic acid sodium salt(DOSS), Sorbitan Monostearate, Sorbitan Tristearate, hexadecyltrimethylammonium bromide (HTAB), Sodium N-lauroylsarcosine, Sodium Oleate,Sodium Myristate, Sodium Stearate, Sodium Palmitate, Gelucire 44/14,ethylenediamine tetraacetic acid (EDTA), Vitamin E d-alpha tocopherylpolyethylene glycol 1000 succinate (TPGS), Lecithin, MW 677-692,Glutanic acid monosodium monohydrate, Labrasol, PEG 8 caprylic/capricglycerides, Transcutol, diethylene glycol monoethyl ether, SolutolHS-15, polyethylene glycol/hydroxystearate, Taurocholic Acid, PluronicF68, Pluronic F108, and Pluronic F127 (or any otherpolyoxyethylene-polyoxypropylene co-polymers (Pluronics®) or saturatedpolyglycolized glycerides (Gelucirs)). Specific example of suchsurfactants that may be used in connection with this invention include,but are not limited to, Span 65, Span 25, Tween 20, Capryol 90, PluronicF108, sodium lauryl sulfate (SLS), Vitamin E TPGS, pluronics andcopolymers. SLS is generally preferred.

The amount of the surfactant (e.g., SLS) relative to the total weight ofthe solid dispersion may be between 0.1-15%. Preferably, it is fromabout 0.5% to about 10%, more preferably from about 05 to about 5%,e.g., about 1%, about 2%, about 3%, about 4%, or about 5%.

In certain embodiments, the amount of the surfactant relative to thetotal weight of the solid dispersion is at least about 0.1, preferablyabout 0.5%. In these embodiments, the surfactant would be present in anamount of no more than about 15%, and preferably no more than about 12%,about 11%, about 10%, about 9%, about 8%, about 7%, about 6%, about 5%,about 4%, about 3%, about 2% or about 1%. An embodiment wherein thesurfactant is in an amount of about 0.5% by weight is preferred.

Candidate surfactants (or other components) can be tested forsuitability for use in the invention in a manner similar to thatdescribed for testing polymers.

Methods of Making Solid Forms of Compound 1

The solid form of Compound 1 can vary depending on the method used toprepare Compound 1. For example, Compound 1 can be prepared using amethod to provide crystalline Compound 1, such as Form A or Form B, orCompound 1 can be prepared using a method to provide amorphous Compound1, for example as a neat preparation or where Compound 1 is a componentin a dispersion such as a solid amorphous dispersion (e.g., a dispersionof Compound 1 and a polymer such as a cellulosic polymer e.g., HPMC orHPMCAS or a pyrrolidone polymer such as PVP/VA).

Form A

Form A of Compound 1 can be prepared, for example, by heating Compound 1to at or above its melting point, for example to about 250° C. and thencooling the compound, thereby providing Compound 1 having a solid stateof Form A. Form A can be characterized by one or more characteristicpeaks as determined using XRPD. For example, Compound 1 as Form A can beidentified by the presence of one or peaks at 20, including one or moreof the following peaks at or about: about 5.0 e.g. 4.99; about 7.8,e.g., 7.75; about 8.5, e.g., 8.46; about 9.2, e.g., 9.21; about 9.9,e.g., 9.92; about 11.9, e.g., 11.93; about 12.6, e.g., 12.64; about13.9, e.g., 13.88; about 14.9, e.g., 14.91; about 15.6, e.g., 15.58;about 16.5, e.g., 16.46; about 18.1, e.g., 18.09; about 18.5, e.g.,18.52; about 21.7, e.g., 20.65; about 22.0, e.g., 21.95; about 23.5,e.g., 23.49; about 25.3, e.g., 25.26; about 28.0, e.g., 28.02; about29.4, e.g., 29.35; and about 30.9, e.g., 30.85.

Form B

Form B of Compound 1 can be prepared, for example, by subjecting aslurry of compound 1 in a solvent to heating and cooling cycles.

In some preferred embodiments, the solvent is a solvent where Compound 1has limited solubility at room temperature, for example, acetone.

The slurry is subject to a plurality of heat/cool cycles, where theslurry is generally warmed to a temperature above room temperature butbelow the boiling point of the solvent, for example about 40° C. toabout 60° C., e.g., about 50° C. The slurry is generally subjected to atleast 2 heat/cool cycles, for example, 2, 3, 4, 5, or 6, preferably 5cycles. Each cycle was timed to last at least about 8 hours (e.g., 4hours of heating followed by 4 hours at room temperature, 6 hours ofheating followed by 6 hours at room temperature, 8 hours of heatingfollowed by 8 hours at room temperature, preferably 12 hours of heatingfollowed by 12 hours at room temperature).

In an alternative embodiment crude Compound 1 can be refluxed in as aslurry in acetonitrile (e.g., 27 volumes of acetonitrile) for 24 hours.The mixture is then cooled, e.g., to about room temperature, e.g., about20° C. Form B is then isolated, for example, by filtration as a white tooff-white. The resulting wet cake is rinsed with acetonitrile (e.g., 5volumes) and dried under vacuum at 50° C. until a constant weight isattained.

Form B can be characterized by one or more characteristic peaks asdetermined using XRPD. For example, Compound 1 as Form B can beidentified by the presence of one or peaks at 20, including one or moreof the following peaks at or about: about 6.2, e.g., 6.17; about 7.6,e.g., 7.61; about 8.4, e.g., 8.40; about 11.0, e.g., 11.02; about 12.3,e.g., 12.33; about 14.8, e.g., 14.83; about 16.1, e.g., 16.14; about17.1, e.g., 17.11; about 18.0, e.g., 17.96; about 18.6, e.g., 18.55;about 19.4, e.g., 19.43; about 21.1, e.g., 21.05; about 22.6, e.g.,22.56; about 23.4, e.g., 23.37; about 23.9, e.g., 23.94; about 24.9,e.g., 24.86; about 25.5, e.g., 25.50; about 26.7, e.g., 26.72; about27.5, e.g., 27.51; about 29.6, e.g., 29.60; about 33.5, e.g., 33.48; andabout 36.8, e.g., 36.78.

Amorphous Compound 1

Amporphous compound 1 can be made using a variety of techniques,including, for example spray drying a solution of Compound 1 to provideamorphous Compound 1, e.g., as a neat solid or as a component of a soliddispersion, said method utilizing spray-drying means to effect saidconversion. For example, Amorphous Compound 1 can be made by convertinga form of Compound 1, e.g., a crystalline form of Compound 1, such asForm A or Form B, into a substantially amorphous form of Compound 1 bydissolving Compound into a solution and spray drying the solution ofCompound 1, thereby converting a form of Compound 1, such as crystallineCompound 1, into amorphous Compound 1. An exemplary process for makingamorphous Compound 1 by converting Form B into a substantially amorphousform of Compound 1 is recited in the examples.

Any method for obtaining amorphous forms of Compound 1, including neatamorphous Compound 1 and solid amorphous dispersions of Compound 1, canbe used including, for example, those described in US 2003/0186952 (seethe documents cited therein at paragraph 1092) and US 2003/0185891). Ingeneral, methods that could be used include those that involve rapidremoval of solvent from a mixture or cooling a molten sample. Suchmethods include, but are not limited to, rotational evaporation,freeze-drying (i.e., lyophilization), vacuum drying, melt congealing,and melt extrusion. However, a preferred embodiment includes amorphousCompound 1, such as a neat preparation or a solid dispersion obtained byspray-drying. Accordingly, in some embodiments, the amorphous productobtained by spray-drying is further dried, for example, to removeresidual solvent.

Preparations disclosed herein, e.g., a pharmaceutical composition, canbe obtained by spray-drying a mixture comprising Compound 1, a suitablepolymer, and an appropriate solvent. Spray drying is a method thatinvolves atomization of a liquid mixture containing, e.g., a solid and asolvent, and removal of the solvent. Atomization can be done, forexample, through a nozzle or on a rotating disk.

Spray drying is a process that converts a liquid feed to a driedparticulate form. Optionally, a secondary drying process such asfluidized bed drying or vacuum drying, may be used to reduce residualsolvents to pharmaceutically acceptable levels. Typically, spray-dryinginvolves contacting a highly dispersed liquid suspension or solution,and a sufficient volume of hot air to produce evaporation and drying ofthe liquid droplets. The preparation to be spray dried can be anysolution, coarse suspension, slurry, colloidal dispersion, or paste thatmay be atomized using the selected spray-drying apparatus. In a standardprocedure, the preparation is sprayed into a current of warm filteredair that evaporates the solvent and conveys the dried product to acollector (e.g., a cyclone). The spent air is then exhausted with thesolvent, or alternatively the spent air is sent to a condenser tocapture and potentially recycle the solvent. Commercially availabletypes of apparatus may be used to conduct the spray-drying. For example,commercial spray dryers are manufactured by Buchi Ltd. and Niro (e.g.,the PSD line of spray driers manufactured by Niro)(see, US 2004/0105820;US 2003/0144257).

Spray-drying typically employs solids loads of material from about 3% toabout 30% by weight, (i.e., drug plus and excipients), for example about4% to about 20% by weight, preferably at least about 10%. In general,the upper limit of solids loads is governed by the viscosity of (e.g.,the ability to pump) the resulting solution and the solubility of thecomponents in the solution. Generally, the viscosity of the solution candetermine the size of the particle in the resulting powder product.

Techniques and methods for spray-drying may be found in Perry's ChemicalEngineering Handbook, 6th Ed., R. H. Perry, D. W. Green & J. O. Maloney,eds.), McGraw-Hill book co. (1984); and Marshall “Atomization andSpray-Drying” 50, Chem. Eng. Prog. Monogr. Series 2 (1954). In general,the spray-drying is conducted with an inlet temperature of from about60° C. to about 200° C., for example, from about 95° C. to about 185°C., from about 110° C. to about 182° C., from about 96° C. to about 108°C., e.g., about 175° C. The spray-drying is generally conducted with anoutlet temperature of from about 30° C. to about 80° C., for examplefrom about 31° C. to about 72° C., about 37° C. to about 41° C. e.g.,about 60° C. The atomization flow rate is generally from about 4 kg/h toabout 12 kg/h, for example, from about 4.3 kg/h to about 10.5 kg/h,e.g., about 6 kg/h or about 10.5 kg/h. The feed flow rate is generallyfrom about 3 kg/h to about 10 kg/h, for example, from about 3.5 kg/h toabout 9.0 kg/h, e.g., about 8 kg/h or about 7.1 kg/h. The atomizationratio is generally from about 0.3 to 1.7, e.g., from about 0.5 to 1.5,e.g., about 0.8 or about 1.5.

Removal of the solvent may require a subsequent drying step, such astray drying, fluid bed drying (e.g., from about room temperature toabout 100° C.), vacuum drying, microwave drying, rotary drum drying orbiconical vacuum drying (e.g., from about room temperature to about 200°C.).

In one embodiment, the solid dispersion is fluid-bed dried.

In preferred processes, the solvent includes a volatile solvent, forexample a solvent having a boiling point of less than about 100° C. Insome embodiments, the solvent includes a mixture of solvents, forexample a mixture of volatile solvents or a mixture of volatile andnon-volatile solvents. Where mixtures of solvents are used, the mixturecan include one or more non-volatile solvents, for example, where thenon-volatile solvent is present in the mixture at less than about 15%,e.g., less than about 12%, less than about 10%, less than about 8%, lessthan about 5%, less than about 3%, or less than about 2%. 10160Preferred solvents are those solvents where Compound 1 has a solubilityof at least about 10 mg/ml (e.g., at least about 15 mg/ml, 20 mg/ml, 25mg/ml, 30 mg/ml, 35 mg/ml, 40 mg/ml, 45 mg/ml, 50 mg/ml, or greater).More preferred solvents include those where Compound 1 has a solubilityof at least about 50 mg/ml.

Exemplary solvents that could be tested include acetone, cyclohexane,dichloromethane, N,N-Dimethylacetamide (DMA), N,N-Dimethylformamide(DMF), 1,3 Dimethyl-2-imidazolidinone (DMI), dimethyl sulfoxide (DMSO),dioxane, ethyl acetate, ethyl ether, glacial acetic acid (HAc), methylethyl ketone (MEK), N-methyl-2-pyrrolidinone (NMP), methyl tert-butylether, tetrahydrofuran (THF) and pentane. Exemplary co-solvents includeacetone/DMSO, acetone/DMF, acetone/water, MEK/water, THF/water,dioxane/water. In a two solvent system, the solvents can be present inof from about 0.1% to about 99.9%. In some preferred embodiments, wateris a co-solvent with acetone where water is present from about 0.1% toabout 15%, for example about 9% to about 11%, e.g., about 10%. In somepreferred embodiments, water is a co-solvent with MEK where water ispresent from about 0.1% to about 15%, for example about 9% to about 11%,e.g., about 10%. In some embodiments the solvent solution include threesolvents. For example, acetone and water can be mixed with a thirdsolvent such as DMA, DMF, DMI, DMSO, or HAc. In instances whereamorphous Compound 1 is a component of a solid amorphous dispersion,preferred solvents dissolve both Compound 1 and the polymer. Suitablesolvents include those described above, for example, MEK, acetone,water, and mixtures thereof.

The particle size and the temperature drying range may be modified toprepare an optimal solid dispersion. As would be appreciated by skilledpractitioners, a small particle size would lead to improved solventremoval. Applicants have found however, that smaller particles can leadto fluffy particles that, under some circumstances do not provideoptimal solid dispersions for downstream processing such as tabletting.At higher temperatures, crystallization or chemical degradation ofCompound 1 may occur. At lower temperatures, a sufficient amount of thesolvent may not be removed. The methods herein provide an optimalparticle size and an optimal drying temperature.

In general, particle size is such that D10 (μm) is less than about 5,e.g., less than about 4.5, less than about 4.0, or less than about 3.5,D50 (μm) is generally less than about 17, e.g., less than about 16, lessthan about 15, less than about 14, less than about 13, and D90 (μm) isgenerally less than about 175, e.g., less than about 170, less thanabout 170, less than about 150, less than about 125, less than about100, less than about 90, less than about 80, less than about 70, lessthan about 60, or less than about less than about 50. In general bulkdensity of the spray dried particles is from about 0.08 g/cc to about0.20 g/cc, e.g., from about 0.10 to about 0.15 g/cc, e.g., about 0.11g/cc or about 0.14 g/cc. Tap density of the spray dried particlesgenerally ranges from about 0.08 g/cc to about 0.20 g/cc, e.g., fromabout 0.10 to about 0.15 g/cc, e.g., about 0.11 g/cc or about 0.14 g/cc,for 10 taps; 0.10 g/cc to about 0.25 g/c, e.g., from about 0.11 to about0.21 g/cc, e.g., about 0.15 g/cc, about 0.19 g/cc, or about 0.21 g/ccfor 500 taps; 0.15 g/cc to about 0.27 g/cc, e.g., from about 0.18 toabout 0.24 g/cc, e.g., about 0.18 g/cc, about 0.19 g/cc, about 0.20 g/c,or about 0.24 g/cc for 1250 taps; and 0.15 g/cc to about 0.27 g/cc,e.g., from about 0.18 to about 0.24 g/cc, e.g., about 0.18 g/c, about0.21 g/cc, about 0.23 g/cc, or about 0.24 g/cc for 2500 taps.

Unless otherwise stated, structures depicted herein are also meant toinclude all isomeric (e.g., enantiomeric, diastereomeric, and geometric(or conformational)) forms of the structure; for example, the R and Sconfigurations for each asymmetric center, (Z) and (E) double bondisomers, and (Z) and (E) conformational isomers. Therefore, singlestereochemical isomers as well as enantiomeric, diastereomeric, andgeometric (or conformational) mixtures of the present compounds arewithin the scope of the invention. All tautomeric forms of the Compound1 are included herein. E.g., Compound 1 may exist as tautomers, both ofwhich are included herein:

Additionally, unless otherwise stated, structures depicted herein arealso meant to include compounds that differ only in the presence of oneor more isotopically enriched atoms. For example, compounds of formula(I), wherein one or more hydrogen atoms are replaced deuterium ortritium, or one or more carbon atoms are replaced by a 13C- or14C-enriched carbon are within the scope of this invention. Suchcompounds are useful, for example, as analytical tools, probes inbiological assays, or compounds with improved therapeutic profile.

Uses, Formulation and Administration

Pharmaceutically Acceptable Compositions

In another aspect of the present invention, pharmaceutically acceptablecompositions are provided, wherein these compositions comprise any ofthe compounds as described herein, and optionally comprise apharmaceutically acceptable carrier, adjuvant or vehicle. In certainembodiments, these compositions optionally further comprise one or moreadditional therapeutic agents.

It will also be appreciated that certain of the compounds of presentinvention can exist in free form for treatment, or where appropriate, asa pharmaceutically acceptable derivative or a prodrug thereof. Accordingto the present invention, a pharmaceutically acceptable derivative or aprodrug includes, but is not limited to, pharmaceutically acceptablesalts, esters, salts of such esters, or any other adduct or derivativewhich upon administration to a patient in need thereof is capable ofproviding, directly or indirectly, a compound as otherwise describedherein, or a metabolite or residue thereof.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgement,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. A“pharmaceutically acceptable salt” means any non-toxic salt or salt ofan ester of a compound of this invention that, upon administration to arecipient, is capable of providing, either directly or indirectly, acompound of this invention or an inhibitorily active metabolite orresidue thereof.

Pharmaceutically acceptable salts are well known in the art. Forexample, S. M. Berge, et al. describe pharmaceutically acceptable saltsin detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporatedherein by reference. Pharmaceutically acceptable salts of the compoundsof this invention include those derived from suitable inorganic andorganic acids and bases. Examples of pharmaceutically acceptable,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid,succinic acid or malonic acid or by using other methods used in the artsuch as ion exchange. Other pharmaceutically acceptable salts includeadipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺(C₁₋₄alkyl)₄ salts. This inventionalso envisions the quaternization of any basic nitrogen-containinggroups of the compounds disclosed herein. Water or oil-soluble ordispersable products may be obtained by such quaternization.Representative alkali or alkaline earth metal salts include sodium,lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, loweralkyl sulfonate and aryl sulfonate.

As described above, the pharmaceutically acceptable compositions of thepresent invention additionally comprise a pharmaceutically acceptablecarrier, adjuvant, or vehicle, which, as used herein, includes any andall solvents, diluents, or other liquid vehicle, dispersion orsuspension aids, surface active agents, isotonic agents, thickening oremulsifying agents, preservatives, solid binders, lubricants and thelike, as suited to the particular dosage form desired. Remington'sPharmaceutical Sciences, Sixteenth Edition, E. W. Martin (MackPublishing Co., Easton, Pa., 1980) discloses various carriers used informulating pharmaceutically acceptable compositions and knowntechniques for the preparation thereof. Except insofar as anyconventional carrier medium is incompatible with the compounds of theinvention, such as by producing any undesirable biological effect orotherwise interacting in a deleterious manner with any othercomponent(s) of the pharmaceutically acceptable composition, its use iscontemplated to be within the scope of this invention. Some examples ofmaterials which can serve as pharmaceutically acceptable carriersinclude, but are not limited to, ion exchangers, alumina, aluminumstearate, lecithin, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, or potassiumsorbate, partial glyceride mixtures of saturated vegetable fatty acids,water, salts or electrolytes, such as protamine sulfate, disodiumhydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zincsalts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, woolfat, sugars such as lactose, glucose and sucrose; starches such as cornstarch and potato starch; cellulose and its derivatives such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; powderedtragacanth; malt; gelatin; talc; excipients such as cocoa butter andsuppository waxes; oils such as peanut oil, cottonseed oil; saffloweroil; sesame oil; olive oil; corn oil and soybean oil; glycols; such apropylene glycol or polyethylene glycol; esters such as ethyl oleate andethyl laurate; agar; buffering agents such as magnesium hydroxide andaluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline;Ringer's solution; ethyl alcohol, and phosphate buffer solutions, aswell as other non-toxic compatible lubricants such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releasingagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator.

Uses of Compounds and Pharmaceutically Acceptable Compositions

In yet another aspect, the present invention provides a method oftreating a condition, disease, or disorder implicated by CFTR. Incertain embodiments, the present invention provides a method of treatinga condition, disease, or disorder implicated by a deficiency of CFTRactivity, the method comprising administering a composition comprising asolid state form of Compound 1 described herein (e.g., Form A, Form B,or amorphous Compound 1, e.g., neat or as a component in a dispersion)to a subject, preferably a mammal, in need thereof.

A “CFTR-mediated disease” as used herein is a disease selected fromcystic fibrosis, Hereditary emphysema, Hereditary hemochromatosis,Coagulation-Fibrinolysis deficiencies, such as Protein C deficiency,Type I hereditary angioedema, Lipid processing deficiencies, such asFamilial hypercholesterolemia, Type 1 chylomicronemia,Abetalipoproteinemia, Lysosomal storage diseases, such as I-celldisease/Pseudo-Hurler, Mucopolysaccharidoses, Sandhof/Tay-Sachs,Crigler-Najjar type I, Polyendocrinopathy/Hyperinsulemia, Diabetesmellitus, Laron dwarfism, Myleoperoxidase deficiency, Primaryhypoparathyroidism, Melanoma, Glycanosis CDG type 1, Hereditaryemphysema, Congenital hyperthyroidism, Osteogenesis imperfecta,Hereditary hypofibrinogenemia, ACT deficiency, Diabetes insipidus (DI),Neurophyseal DI, Neprogenic DI, Charcot-Marie Tooth syndrome,Perlizaeus-Merzbacher disease, neurodegenerative diseases such asAlzheimer's disease, Parkinson's disease, Amyotrophic lateral sclerosis,Progressive supranuclear plasy, Pick's disease, several polyglutamineneurological disorders asuch as Huntington, Spinocerebullar ataxia typeI, Spinal and bulbar muscular atrophy, Dentatorubal pallidoluysian, andMyotonic dystrophy, as well as Spongiform encephalopathies, such asHereditary Creutzfeldt-Jakob disease, Fabry disease,Straussler-Scheinker syndrome, COPD, dry-eye disease, and Sjogren'sdisease.

In certain embodiments, the present invention provides a method oftreating cystic fibrosis, hereditary emphysema, hereditaryhemochromatosis, coagulation-fibrinolysis deficiencies, such as proteinC deficiency, Type I hereditary angioedema, lipid processingdeficiencies, such as familial hypercholesterolemia, Type 1chylomicronemia, abetalipoproteinemia, lysosomal storage diseases, suchas I-cell disease/pseudo-Hurler, mucopolysaccharidoses,Sandhof/Tay-Sachs, Crigler-Najjar type II,polyendocrinopathy/hyperinsulemia, Diabetes mellitus, Laron dwarfism,myleoperoxidase deficiency, primary hypoparathyroidism, melanoma,glycanosis CDG type 1, congenital hyperthyroidism, osteogenesisimperfecta, hereditary hypofibrinogenemia, ACT deficiency, Diabetesinsipidus (DI), neurophyseal DI, neprogenic DI, Charcot-Marie Toothsyndrome, Perlizaeus-Merzbacher disease, neurodegenerative diseases suchas Alzheimer's disease, Parkinson's disease, amyotrophic lateralsclerosis, progressive supranuclear plasy, Pick's disease, severalpolyglutamine neurological disorders asuch as Huntington,spinocerebullar ataxia type I, spinal and bulbar muscular atrophy,dentatorubal pallidoluysian, and myotonic dystrophy, as well asspongiform encephalopathies, such as hereditary Creutzfeldt-Jakobdisease (due to prion protein processing defect), Fabry disease,Straussler-Scheinker syndrome, COPD, dry-eye disease, or Sjogren'sdisease, comprising the step of administering to said mammal aneffective amount of a composition comprising a solid state form ofCompound 1 described herein (e.g., Form A, or Form B, or amorphousCompound 1, e.g., neat or as a component in a dispersion).

According to an alternative preferred embodiment, the present inventionprovides a method of treating cystic fibrosis comprising the step ofadministering to said mammal a composition comprising a solid state formof Compound 1 described herein (e.g., Form A, or Form B, or amorphousCompound 1, e.g., neat or as a component in a dispersion).

According to the invention an “effective amount” of a solid state formof Compound 1 (e.g., Form A, or Form B, or amorphous Compound 1, e.g.,neat or as a component in a dispersion) or a pharmaceutically acceptablecomposition thereof is that amount effective for treating or lesseningthe severity of any of the diseases recited above.

A solid state form of Compound 1 (e.g., Form A, or Form B, or amorphousCompound 1, e.g., neat or as a component in a dispersion) or apharmaceutically acceptable composition thereof may be administeredusing any amount and any route of administration effective for treatingor lessening the severity of one or more of the diseases reicted above.

In certain embodiments, a solid state form of Compound 1 describedherein (e.g., Form A, or Form B, or amorphous Compound 1, e.g., neat oras a component in a dispersion) or a pharmaceutically acceptablecomposition thereof is useful for treating or lessening the severity ofcystic fibrosis in patients who exhibit residual CFTR activity in theapical membrane of respiratory and non-respiratory epithelia. Thepresence of residual CFTR activity at the epithelial surface can bereadily detected using methods known in the art, e.g., standardelectrophysiological, biochemical, or histochemical techniques. Suchmethods identify CFTR activity using in vivo or ex vivoelectrophysiological techniques, measurement of sweat or salivary Cl⁻concentrations, or ex vivo biochemical or histochemical techniques tomonitor cell surface density. Using such methods, residual CFTR activitycan be readily detected in patients heterozygous or homozygous for avariety of different mutations, including patients homozygous orheterozygous for the most common mutation, ΔF508.

In one embodiment, a solid state form of Compound 1 described herein(e.g., Form A, or Form B, or amorphous Compound 1, e.g., neat or as acomponent in a dispersion) or a pharmaceutically acceptable compositionthereof is useful for treating or lessening the severity of cysticfibrosis in patients within certain genotypes exhibiting residual CFTRactivity, e.g., class III mutations (impaired regulation or gating),class IV mutations (altered conductance), or class V mutations (reducedsynthesis)(Lee R. Choo-Kang, Pamela L., Zeitlin, Type I, IIII, IV, and Vcystic fibrosis Tansmembrane Conductance Regulator Defects andOpportunities of Therapy; Current Opinion in Pulmonary Medicine6:521-529, 2000). Other patient genotypes that exhibit residual CFTRactivity include patients homozygous for one of these classes orheterozygous with any other class of mutations, including class Imutations, class I mutations, or a mutation that lacks classification.

In one embodiment, a solid state form of Compound 1 described herein(e.g., Form A, or Form B, or amorphous Compound 1, e.g., neat or as acomponent in a dispersion) or a pharmaceutically acceptable compositionthereof is useful for treating or lessening the severity of cysticfibrosis in patients within certain clinical phenotypes, e.g., amoderate to mild clinical phenotype that typically correlates with theamount of residual CFTR activity in the apical membrane of epithelia.Such phenotypes include patients exhibiting pancreatic insufficiency orpatients diagnosed with idiopathic pancreatitis and congenital bilateralabsence of the vas deferens, or mild lung disease.

The exact amount required will vary from subject to subject, dependingon the species, age, and general condition of the subject, the severityof the infection, the particular agent, its mode of administration, andthe like. The compounds of the invention are preferably formulated indosage unit form for ease of administration and uniformity of dosage.The expression “dosage unit form” as used herein refers to a physicallydiscrete unit of agent appropriate for the patient to be treated. Itwill be understood, however, that the total daily usage of the compoundsand compositions of the present invention will be decided by theattending physician within the scope of sound medical judgment. Thespecific effective dose level for any particular patient or organismwill depend upon a variety of factors including the disorder beingtreated and the severity of the disorder; the activity of the specificcompound employed; the specific composition employed; the age, bodyweight, general health, sex and diet of the patient; the time ofadministration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed, andlike factors well known in the medical arts. The term “patient”, as usedherein, means an animal, preferably a mammal, and most preferably ahuman.

The pharmaceutically acceptable compositions of this invention can beadministered to humans and other animals orally, rectally, parenterally,intracistemally, intravaginally, intraperitoneally, topically (as bypowders, ointments, or drops), bucally, as an oral or nasal spray, orthe like, depending on the severity of the infection being treated. Incertain embodiments, the compounds of the invention may be administeredorally or parenterally at dosage levels of about 0.01 mg/kg to about 50mg/kg and preferably from about 1 mg/kg to about 25 mg/kg, of subjectbody weight per day, one or more times a day, to obtain the desiredtherapeutic effect.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformatnide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a compound of the present invention,it is often desirable to slow the absorption of the compound fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material withpoor water solubility. The rate of absorption of the compound thendepends upon its rate of dissolution that, in turn, may depend uponcrystal size and crystalline form. Alternatively, delayed absorption ofa parenterally administered compound form is accomplished by dissolvingor suspending the compound in an oil vehicle. Injectable depot forms aremade by forming microencapsule matrices of the compound in biodegradablepolymers such as polylactide-polyglycolide. Depending upon the ratio ofcompound to polymer and the nature of the particular polymer employed,the rate of compound release can be controlled. Examples of otherbiodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping thecompound in liposomes or microemulsions that are compatible with bodytissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polethylene glycols and the like.

The active compounds can also be in microencapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, eardrops, and eye drops are also contemplated asbeing within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms are prepared by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

It will also be appreciated that the solid state form of Compound 1described herein (e.g., Form A, or Form B, or amorphous Compound 1,e.g., neat or as a component in a dispersion) or a pharmaceuticallyacceptable composition thereof can be employed in combination therapies,that is, Form A or Form B or a pharmaceutically acceptable compositionthereof can be administered concurrently with, prior to, or subsequentto, one or more other desired therapeutics or medical procedures. Theparticular combination of therapies (therapeutics or procedures) toemploy in a combination regimen will take into account compatibility ofthe desired therapeutics and/or procedures and the desired therapeuticeffect to be achieved. It will also be appreciated that the therapiesemployed may achieve a desired effect for the same disorder (forexample, an inventive compound may be administered concurrently withanother agent used to treat the same disorder), or they may achievedifferent effects (e.g., control of any adverse effects). As usedherein, additional therapeutic agents that are normally administered totreat or prevent a particular disease, or condition, are known as“appropriate for the disease, or condition, being treated”.

In one embodiment, the additional agent is selected from a mucolyticagent, bronchodialator, an anti-biotic, an anti-infective agent, ananti-inflammatory agent, a CFTR modulator other than a compound of thepresent invention, or a nutritional agent.

The amount of additional therapeutic agent present in the compositionsof this invention will be no more than the amount that would normally beadministered in a composition comprising that therapeutic agent as theonly active agent. Preferably the amount of additional therapeutic agentin the presently disclosed compositions will range from about 50% to100% of the amount normally present in a composition comprising thatagent as the only therapeutically active agent.

A solid state form of Compound 1 described herein (e.g., Form A, or FormB, or amorphous Compound 1, e.g., neat or as a component in adispersion) or a pharmaceutically acceptable composition thereof mayalso be incorporated into compositions for coating an implantablemedical device, such as prostheses, artificial valves, vascular grafts,stents and catheters. Accordingly, the present invention, in anotheraspect, includes a composition for coating an implantable devicecomprising a solid state form of Compound 1 described herein (e.g., FormA, or Form B, or amorphous Compound 1, e.g., neat or as a component in adispersion) or a pharmaceutically acceptable composition thereof, and inclasses and subclasses herein, and a carrier suitable for coating saidimplantable device. In still another aspect, the present inventionincludes an implantable device coated with a composition comprising asolid state form of Compound 1 described herein (e.g., Form A, or FormB, or amorphous Compound 1, e.g., neat or as a component in adispersion) or a pharmaceutically acceptable composition thereof, and acarrier suitable for coating said implantable device. Suitable coatingsand the general preparation of coated implantable devices are describedin U.S. Pat. Nos. 6,099,562; 5,886,026; and 5,304,121. The coatings aretypically biocompatible polymeric materials such as a hydrogel polymer,polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylacticacid, ethylene vinyl acetate, and mixtures thereof. The coatings mayoptionally be further covered by a suitable topcoat of fluorosilicone,polysaccarides, polyethylene glycol, phospholipids or combinationsthereof to impart controlled release characteristics in the composition.

In order that the invention described herein may be more fullyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner.

EXAMPLES

Methods & Materials

Differential Scanning Calorimetry (DSC)

The Differential scanning calorimetry (DSC) data of Form A, Form B, andamorphous Compound 1 were collected using a DSC Q100 V9.6 Build 290 (TAInstruments, New Castle, Del.). Temperature was calibrated with indiumand heat capacity was calibrated with sapphire. Samples of 3-6 mg wereweighed into aluminum pans that were crimped using lids with 1 pin hole.The samples were scanned from 25° C. to 350° C. at a heating rate of 10°C./min and with a nitrogen gas purge of 50 mil/min. Data were collectedby Thermal Advantage Q Series™ version 2.2.0.248 software and analyzedby Universal Analysis software version 4.1D (TA Instruments, New Castle,Del.). The reported numbers represent single analyses.

Thermogravimetric Analysis (TGA)

Thermal gravimetric analysis (TGA) was performed with a TGA Q500 V6.3Build 189 (TA Instruments, New Castle, Del.) was used for TGAmeasurement. Temperature was equilibrated by Curie point with nickel.Samples of 10-20 mg were scanned from 25° C. to 350° C. at a heatingrate of 10° C./min. A nitrogen gas balance purge of 10 ml/min and asample purge of 90 ml/min were used. Data were collected by ThermalAdvantage Q Series™ software version 2.2.0.248 and analyzed by UniversalAnalysis software version 4.1D (TA Instruments, New Castle, Del.). Thereported numbers represent single analyses.

XRPD (X-ray Powder Diffraction)

The X-Ray diffraction (XRD) data of Form A, Form B, and amorphousCompound 1 were collected on a Bruker D8 DISCOVER with GADDS powderdiffractometer with HI-STAR 2-dimensional detector and a flat graphitemonochromator. Cu sealed tube with Kα radiation was used at 40 kV, 35mA. The samples were placed on zero-background silicon wafers at 25° C.For each sample, two data frames were collected at 120 seconds each at 2different 20 angles: 8° and 26°. The frames data were integrated withGADDS software and merged with DIFFRACT^(plus)EVA software.

Synthesis ofN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide(Compound 1)

2-Phenylaminomethylene-malonic acid diethyl ester

A mixture of aniline (25.6 g, 0.275 mol) and diethyl2-(ethoxymethylene)malonate (62.4 g, 0.288 mol) was heated at 140-150°C. for 2 h. The mixture was cooled to room temperature and dried underreduced pressure to afford 2-phenylaminomethylene-malonic acid diethylester as a solid, which was used in the next step without furtherpurification. ¹H NMR (DMSO-d₆) δ 11.00 (d, 1H), 8.54 (d, J=13.6 Hz, 1H),7.36-7.39 (m, 2H), 7.13-7.17 (m, 3H), 4.174.33 (m, 4H), 1.18-1.40 (m,6H).

4-Hydroxyquinoline-3-carboxylic acid ethyl ester

A 1 L three-necked flask fitted with a mechanical stirrer was chargedwith 2-phenylaminomethylene-malonic acid diethyl ester (26.3 g, 0.100mol), polyphosphoric acid (270 g) and phosphoryl chloride (750 g). Themixture was heated to 70° C. and stirred for 4 h. The mixture was cooledto room temperature and filtered. The residue was treated with aqueousNa₂CO₃ solution, filtered, washed with water and dried.4-Hydroxyquinoline-3-carboxylic acid ethyl ester was obtained as a palebrown solid (15.2 g, 70%). The crude product was used in next stepwithout further purification.

4-Oxo-1,4-dihydroquinoline-3-carboxylic acid

4-Hydroxyquinoline-3-carboxylic acid ethyl ester (15 g, 69 mmol) wassuspended in sodium hydroxide solution (2N, 150 mL) and stirred for 2 hat reflux. After cooling, the mixture was filtered, and the filtrate wasacidified to pH 4 with 2N HCl. The resulting precipitate was collectedvia filtration, washed with water and dried under vacuum to give4-oxo-1,4-dihydroquinoline-3-carboxylic acid as a pale white solid (10.5g, 92%). ¹H NMR (DMSO-d₆) δ 15.34 (s, 1H), 13.42 (s, 1H), 8.89 (s, 1H),8.28 (d, J=8.0 Hz, 1H), 7.88 (m, 1H), 7.81 (d, J=8.4 Hz, 1H), 7.60 (m,1H).

Carbonic acid 2,4-di-tert-butyl-phenyl ester methyl ester

Methyl chloroformate (58 mL, 750 mmol) was added dropwise to a solutionof 2,4-di-tert-butyl-phenol (103.2 g, 500 mmol), Et₃N (139 mL, 1000mmol) and DMAP (3.05 g, 25 mmol) in dichloromethane (400 mL) cooled inan ice-water bath to 0° C. The mixture was allowed to warm to roomtemperature while stirring overnight, then filtered through silica gel(approx. 1 L) using 10% ethyl acetate-hexanes (˜4 L) as the eluent. Thecombined filtrates were concentrated to yield carbonic acid2,4-di-tert-butyl-phenyl ester methyl ester as a yellow oil (132 g,quant.). ¹H NMR (400 MHz, DMSO-d₆) δ 7.35 (d, J=2.4 Hz, 1H), 7.29 (dd,J=8.5, 2.4 Hz, 1H), 7.06 (d, J=8.4 Hz, 1H), 3.85 (s, 3H), 1.30 (s, 9H),1.29 (s, 9H).

Carbonic acid 2,4-di-tert-butyl-5-nitro-phenyl ester methyl ester andCarbonic acid 2,4-di-tert-butyl-6-nitro-phenyl ester methyl ester

To a stirring mixture of carbonic acid 2,4-di-tert-butyl-phenyl estermethyl ester (4.76 g, 180 mmol) in conc. sulfuric acid (2 mL), cooled inan ice-water bath, was added a cooled mixture of sulfuric acid (2 mL)and nitric acid (2 mL). The addition was done slowly so that thereaction temperature did not exceed 50° C. The reaction was allowed tostir for 2 h while warming to room temperature. The reaction mixture wasthen added to ice-water and extracted into diethyl ether. The etherlayer was dried (MgSO₄), concentrated and purified by columnchromatography (0-10% ethyl acetate-hexanes) to yield a mixture ofcarbonic acid 2,4-di-tert-butyl-5-nitro-phenyl ester methyl ester andcarbonic acid 2,4-di-tert-butyl-6-nitro-phenyl ester methyl ester as apale yellow solid (4.28 g), which was used directly in the next step.

2,4-Di-tert-butyl-5-nitro-phenol and 2,4-Di-tert-butyl-6-nitro-phenol

The mixture of carbonic acid 2,4-di-tert-butyl-5-nitro-phenyl estermethyl ester and carbonic acid 2,4-di-tert-butyl-6-nitro-phenyl estermethyl ester (4.2 g, 14.0 mmol) was dissolved in MeOH (65 mL) before KOH(2.0 g, 36 mmol) was added. The mixture was stirred at room temperaturefor 2 h. The reaction mixture was then made acidic (pH 2-3) by addingconc. HCl and partitioned between water and diethyl ether. The etherlayer was dried (MgSO₄), concentrated and purified by columnchromatography (0-5% ethyl acetate-hexanes) to provide2,4-di-tert-butyl-5-nitro-phenol (1.31 g, 29% over 2 steps) and2,4-di-tert-butyl-6-nitro-phenol. 2,4-Di-tert-butyl-5-nitro-phenol: ¹HNMR (400 MHz, DMSO-d₆) δ 10.14 (s, 1H, OH), 7.34 (s, 1H), 6.83 (s, 1H),1.36 (s, 9H), 1.30 (s, 9H). 2,4-Di-tert-butyl-6-nitro-phenol: ¹H NMR(400 MHz, CDCl₃) δ 11.48 (s, 1H), 7.98 (d, J=2.5 Hz, 1H), 7.66 (d, J=2.4Hz, 1H), 1.47 (s, 9H), 1.34 (s, 9H).

5-Amino-2,4-di-tert-butyl-phenol

To a reluxing solution of 2,4-di-tert-butyl-5-nitro-phenol (1.86 g, 7.40mmol) and ammonium formate (1.86 g) in ethanol (75 mL) was added Pd-5%wt. on activated carbon (900 mg). The reaction mixture was stirred atreflux for 2 h, cooled to room temperature and filtered through Celite.The Celite was washed with methanol and the combined filtrates wereconcentrated to yield 5-amino-2,4-di-tert-butyl-phenol as a grey solid(1.66 g, quant.). ¹H NMR (400 MHz, DMSO-d₆) δ 8.64 (s, 1H, OH), 6.84 (s,1H), 6.08 (s, 1H), 4.39 (s, 2H, NH₂), 1.27 (m, 18H); HPLC ret. time 2.72min, 10-99% CH₃CN, 5 min run; ESI-MS 222.4 m/z [M+H]+.

N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoqunoline-3-carboxamide

To a suspension of 4-oxo-1,4-dihydroquinolin-3-carboxylic acid (35.5 g,188 mmol) and HBTU (85.7 g, 226 mmol) in DMF (280 mL) was added Et₃N(63.0 mL, 451 mmol) at ambient temperature. The mixture becamehomogeneous and was allowed to stir for 10 min before5-amino-2,4-di-tert-butyl-phenol (50.0 g, 226 mmol) was added in smallportions. The mixture was allowed to stir overnight at ambienttemperature. The mixture became heterogeneous over the course of thereaction. After all of the acid was consumed (LC-MS analysis, MH+190,1.71 min), the solvent was removed in vacuo. EtOH was added to theorange solid material to produce a slurry. The mixture was stirred on arotovap (bath temperature 65° C.) for 15 min without placing the systemunder vacuum. The mixture was filtered and the captured solid was washedwith hexanes to provide a white solid that was the EtOH crystalate. Et₂Owas added to the material obtained above until a slurry was formed. Themixture was stirred on a rotovapor (bath temperature 25° C.) for 15 minwithout placing the system under vacuum. The mixture was filtered andthe solid captured. This procedure was performed a total of five times.The solid obtained after the fifth precipitation was placed under vacuumovernight to provide 8N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideas a white powdery solid (38 g, 52%).

HPLC ret. time 3.45 min, 10-99% CH₃CN, 5 min run; ¹H NMR (400 MHz,DMSO-d₆) δ 12.88 (s, 1H), 11.83 (s, 1H), 9.20 (s, 1H), 8.87 (s, 1H),8.33 (dd, J=8.2, 1.0 Hz, 1H), 7.83-7.79 (m, 1H), 7.76 (d, J=7.7 Hz, 1H),7.54-7.50 (m, 1H), 7.17 (s, 1H), 7.10 (s, 1H), 1.38 (s, 9H), 1.37 (s,9H); ESI-MS 393.3 m/z [M+H]*.

Set forth below is the characterizing data for Compound 1:

TABLE 2 LC-MS LC-RT Cmd No. M + 1 min 1 393.2 3.71

The XRPD spectrum of Compound 1 is shown in FIG. 1.

¹H NMR data for Compound 1 in shown in FIG. 2. The DSC trace of Compound1 is shown in FIG. 3.

Preparation of Form A

Form A was obtained by heating Compound 1 as a solid to 250° C. andcooling to room temperature. The DSC thermogram on Compound 1 (See FIG.6) shows that the compound undergoes a melt with an onset temperature of195° C., followed by a re-crystallisation with onset at 220° C.

The XRPD spectrum of Form A is shown in FIG. 4.

The DSC data for Form A is shown in FIG. 5.

The TGA trace for Form A is shown in FIG. 6.

Preparation of Form B

Crude Compound 1 was a slurry in refluxing acetonitrile (27 volumes) for24 hours. After 24 hours, the mixture was allowed to cool to 20° C. FormB was isolated by filtration as a white to off-white. The wet cake wasrinsed with acetonitrile (5 volumes) and dried under vacuum at 50° C.until a constant weight is attained, thereby providing Form B.

The XRPD spectrum of Form B is shown in FIG. 7.

The DSC trace of Form B is shown in FIG. 8.

The TGA trace for Form B is shown in FIG. 9.

Single crystal data was obtained for Form B, providing detail additionaldetail about the crystal structure, including lattice size and packing.

Crystal preparation:

1 g of Compound 1 was added to 10 ml of isopropanol acetate. Thesuspension was heated and remained at 60° C. for 3 hours. The suspensionwas cooled to room temperature and remained stirred overnight. Thesuspended solid was filtered and washed with isopropanol acetate. Thecollected solid was dried under vacuum at room temperature. 300 mg ofthe dried solid was dissolved in 5 ml of 10% aqueous ethyl acetatesolution. The solution was heated to 70° C. for 10 minutes before it wascooled to room temperature. Over time, crystals grew in the vial.

Experimental

A single crystal of Form B was mounted on a MicroMount loop and centeredon a Bruker Apex II diffractometer that was equipped with a sealedcopper X-ray tube and Apex II CCD detector. Initially, 3 sets of 40frames were collected to determine a preliminary unit cell. Subsequentlya full data set consisting of 15 scans and 6084 frames was acquired.Data collection was performed at 100 K. Data were integrated and scaledusing Apex II software from Bruker AXS. Integration and scaling resultedin 7528 reflections, 3071 of which were unique [R(int))=0.0466],Structure was solved by direct methods in space group P2₁ using SHELXTLsoftware. Refinement was performed with full-matrix least-square methodon F² using SHELXTL software as well. Altogether 375 parameters wereused in refinement resulting in reflection to parameter ratio of 8.19.The final refinement afforded a chiral structure with a Flack parameterof 0.0(3). The final refinement index was wR2=0.1242 and R1=0.0663(wR2=0.1137 and R1=0.0482 for reflections with I>2 sigma (I)).

A conformational picture of Form B is provided in FIG. 10, which is incolor.

TABLE 1 Crystal data and structure refinement for Compound 1.Identification code Compound 1 Empirical formula C24 H28 N2 O3 Formulaweight 392.48 Temperature 100(2) K Wavelength 1.54178 Å Crystal systemmonoclinic Space group P2_(t) Unit cell dimensions a = 11.8011(7) Å α =90°. b = 5.9819(3) Å β = 105.110(4)°. c = 14.7974(8) Å γ = 90°. Volume1008.48(10) Å³ Z 2 Density (calculated) 1.293 Mg/m³ Absorptioncoefficient 0.681 mm⁻¹ F(000) 420 Crystal size 0.20 × 0.08 × 0.08 mm³Theta range for data collection 3.09 to 68.67°. Index ranges −14 <= h <=14, −7 <= k <= 7, −14 <= l <= 17 Reflections collected 7528 Independentreflections 3071 [R(int) = 0.0466] Completeness to theta = 68.67° 94.6%Max. and min. transmission 0.9475 and 0.8758 Refinement methodFull-matrix least-squares on F² Data/restraints/parameters 3071/1/375Goodness-of-fit on F² 1.001 Final R indices [I>2sigma(I)] R1 = 0.0482,wR2 = 0.1137 R indices (all data) R1 = 0.0663, wR2 = 0.1242 Absolutestructure parameter 0.0(3) Extinction coefficient 0.0008(6) Largestdiff. peak and hole 0.200 and −0.218 e.Å⁻³

TABLE 2 Atomic coordinates (×10⁴ and equivalent isotropic displacementparameters (Å² ×10³) for Compound 1. U(eq) is defined as one third ofthe trace of the orthogonalized U^(ij) tensor. x y z U(eq) N(1)11624(2)  −530(5) 8667(2) 33(1) C(3) 10407(3)  2650(6) 8133(2) 31(1)C(4) 10335(3)  2115(6) 7158(2) 30(1) C(5) 11006(3)   150(6) 7013(2)31(1) C(2) 11034(3)  1269(6) 8830(2) 32(1) C(8) 12233(3)  −3643(7) 6701(3) 38(1) C(7) 11610(3)  −2371(7)  5947(3) 37(1) C(6) 11015(3) −530(6) 6093(2) 35(1) C(9) 12255(3)  −3039(7)  7618(2) 36(1) C(11)9786(3) 4549(6) 8431(2) 32(1) C(15) 6468(3) 9414(6) 7354(2) 30(1) C(19)7049(3) 7025(6) 6127(2) 31(1) C(18) 8461(3) 8580(6) 8706(2) 32(1) C(13)8275(3) 7448(6) 7859(2) 29(1) C(16) 6578(3) 10442(6)  8223(2) 31(1)C(17) 7639(3) 10044(6)  8889(2) 30(1) C(14) 7271(3) 7965(6) 7130(2)30(1) C(23) 5586(3) 11841(6)  8438(2) 34(1) C(21) 8075(3) 7722(7)5705(2) 35(1) C(22) 5943(3) 8034(7) 5474(2) 35(1) C(20) 6879(3) 4481(7)6096(3) 37(1) C(24) 4478(3) 11888(7)  7605(3) 39(1) C(25) 5981(3)14254(7)  8672(3) 39(1) C(26) 5207(3) 10760(7)  9264(3) 37(1) N(12)9082(2) 5775(5) 7752(2) 31(1) O(11) 9923(2) 4910(4) 9289(2) 37(1) O(4)9748(2) 3206(4) 6485(2) 35(1) O(17) 7888(2) 11078(5)  9758(2) 37(1)C(10) 11644(3)  −1165(6)  7761(2) 32(1)

TABLE 3 Bond lengths [Å] and angles [°] for Compound 1. Length AngleAngle Bond (Å) Bond (deg) Bond (deg) N(1)—C(2) 1.337(5) C(2)—N(1)—C(10)122.0(3) C(15)—C(16)—C(23) 122.4(3) N(1)—C(10) 1.400(5) C(2)—C(3)—C(4)119.4(3) C(18)—C(17)—O(17) 118.2(3) C(3)—C(2) 1.377(5) C(2)—C(3)—C(11)116.6(3) C(18)—C(17)—C(16) 120.9(3) C(3)—C(4) 1.458(4) C(4)—C(3)—C(11)123.9(3) O(17)—C(17)—C(16) 120.8(3) C(3)—C(11) 1.481(5) O(4)—C(4)—C(3)123.8(3) C(15)—C(14)—C(13) 116.3(3) C(4)—O(4) 1.240(4) O(4)—C(4)—C(5)120.9(3) C(15)—C(14)—C(19) 120.1(3) C(4)—C(5) 1.465(5) C(3)—C(4)—C(5)115.3(3) C(13)—C(14)—C(19) 123.5(3) C(5)—C(10) 1.406(5) C(10)—C(5)—C(6)117.2(3) C(25)—C(23)—C(16) 110.9(3) C(5)—C(6) 1.423(5) C(10)—C(5)—C(4)122.2(3) C(25)—C(23)—C(24) 107.9(3) C(8)—C(7) 1.391(5) C(6)—C(5)—C(4)120.6(3) C(16)—C(23)—C(24) 112.1(3) C(8)—C(9) 1.398(5) N(1)—C(2)—C(3)123.4(3) C(25)—C(23)—C(26) 110.5(3) C(7)—C(6) 1.353(5) C(7)—C(8)—C(9)120.6(4) C(16)—C(23)—C(26) 109.2(3) C(9)—C(10) 1.379(5) C(6)—C(7)—C(8)120.3(4) C(24)—C(23)—C(26) 106.1(3) C(11)—O(11) 1.255(4) C(7)—C(6)—C(5)121.2(3) C(11)—N(12)—C(13) 127.2(3) C(11)—N(12) 1.343(4) C(10)—C(9)—C(8)118.7(3) C(9)—C(10)—N(1) 120.5(3) C(15)—C(14) 1.387(5) O(11)—C(11)—N(12)123.7(3) C(9)—C(10)—C(5) 121.9(3) C(15)—C(16) 1.399(5) O(11)—C(11)—C(3)119.4(3) N(1)—C(10)—C(5) 117.6(3) C(19)—C(22) 1.531(4) N(12)—C(11)—C(3)117.0(3) C(19)—C(20) 1.534(5) C(14)—C(15)—C(16) 126.1(3) C(19)—C(14)1.544(4) C(22)—C(19)—C(20) 106.8(3) C(19)—C(21) 1.558(5)C(22)—C(19)—C(14) 111.5(3) C(18)—C(17) 1.385(5) C(20)—C(19)—C(14)112.2(3) C(18)—C(13) 1.390(5) C(22)—C(19)—C(21) 105.4(3) C(13)—C(14)1.413(4) C(20)—C(19)—C(21) 111.3(3) C(13)—N(12) 1.418(4)C(14)—C(19)—C(21) 109.4(3) C(16)—C(17) 1.397(4) C(17)—C(18)—C(13)122.0(3) C(16)—C(23) 1.539(5) C(18)—C(13)—C(14) 119.0(3) C(17)—O(17)1.387(4) C(18)—C(13)—N(12) 119.4(3) C(23)—C(25) 1.528(5)C(14)—C(13)—N(12) 121.5(3) C(23)—C(24) 1.546(5) C(17)—C(16)—C(15)115.1(3) C(23)—C(26) 1.548(5) C(17)—C(16)—C(23) 122.4(3)

Symmetry transformations used to generate equivalent atoms:

TABLE 4 Anisotropic displacement parameters (Å² × 10³)for Compound 1.The anisotropic displacement factor exponent takes the form:−2π²[h²a*²U¹¹ + . . . + 2 h k a* b* U¹²] U¹¹ U²² U³³ U²³ U¹³ U¹² N(1)42(1) 41(2) 14(2) 5(1) 4(1) 3(1) C(3) 34(2) 40(2) 16(2) −1(1)  4(1)−4(1)  C(4) 34(2) 38(2) 17(2) 0(1) 4(1) −1(1)  C(5) 34(2) 42(2) 17(2)−2(1)  6(1) −6(1)  C(2) 37(2) 42(2) 16(2) 1(1) 5(1) 1(2) C(8) 44(2)41(2) 30(2) −4(2)  10(1)  5(2) C(7) 46(2) 44(2) 22(2) −4(1)  9(1) −5(2) C(6) 41(2) 40(2) 23(2) 1(2) 9(1) −1(2)  C(9) 41(2) 40(2) 24(2) 5(1) 4(1)3(2) C(11) 35(2) 41(2) 18(2) 1(1) 4(1) −4(2)  C(15) 37(2) 37(2) 15(2)4(1) 3(1) 1(1) C(19) 38(2) 38(2) 14(2) 2(1) 5(1) 4(1) C(18) 36(2) 42(2)14(2) 4(1) 0(1) 0(1) C(13) 39(2) 34(2) 16(2) 2(1) 9(1) −3(1)  C(16)46(2) 29(2) 19(2) 1(1) 10(1)  −3(1)  C(17) 43(2) 33(2) 14(2) −2(1)  7(1)−6(1)  C(14) 38(2) 38(2) 11(2) 1(1) 3(1) −3(2)  C(23) 46(2) 40(2) 20(2)2(1) 13(1)  3(2) C(21) 51(2) 45(2)  8(2) 2(1) 7(1) 0(2) C(22) 44(2)41(2) 16(2) −7(1)  1(1) 2(2) C(20) 40(2) 46(2) 20(2) −1(2)  3(1) 1(2)C(24) 44(2) 49(2) 24(2) −2(2)  10(1)  5(2) C(25) 52(2) 43(2) 24(2) 3(1)12(2)  9(2) C(26) 48(2) 40(2) 24(2) 1(1) 14(1)  0(2) N(12) 40(1) 41(2)12(2) 0(1) 5(1) 0(1) O(11) 48(1) 47(1) 13(1) 1(1) 4(1) 5(1) O(4) 46(1)46(2) 12(1) 3(1) 4(1) 7(1) O(17) 44(1) 45(2) 18(1) −6(1)  4(1) 4(1)C(10) 37(2) 37(2) 21(2) 0(1) 8(1) −2(2) 

TABLE 5 Hydrogen coordinates (×10⁴) and isotropic displacementparameters (Å² ×10³) for Compound 1. x y z U(eq) H(7) 11560(30) −2840(70)  5320(30) 35(10) H(9) 12670(30)  −3980(70)  8120(30) 38(10)H(8) 12680(30)  −4860(70)  6660(30) 36(10) H(6) 10550(30)   350(80)5580(30) 51(13) H(15) 5770(30) 9730(70) 6900(30) 30(9)  H(18) 9150(20)8310(50) 9160(20) 12(7)  H(17) 8620(30) 10600(60)  10030(30)  25(9) H(20A) 7470(30) 3650(70) 6460(30) 32(10) H(20B) 6130(30) 4320(80)6280(30) 43(11) H(21A) 8840(30) 6840(70) 5980(30) 40(11) H(21B) 8160(30)9370(80) 5790(30) 42(11) H(22B) 5990(30) 9620(70) 5480(30) 31(9)  H(22A)5790(30) 7200(80) 4820(30) 48(12) H(24A) 3800(40) 12810(90)  7750(30)57(13) H(24B) 4210(30) 10410(70)  7420(30) 34(10) H(25A) 5370(30)15130(60)  8770(20) 24(9)  H(25B) 6240(30) 15040(70)  8150(30) 41(11)H(25C) 6690(30) 14100(80)  9230(30) 44(11) H(26A) 4600(30) 11790(60) 9320(20) 17(8)  H(26B) 5000(30) 9350(70) 9090(30) 28(9)  H(1) 12000(30) −1450(70)  9140(30) 40(11) H(2) 11050(40)  1550(80) 9460(40) 56(13)H(26C) 5950(30) 10770(80)  9820(30) 51(12) H(24C) 4720(40) 12850(100)7170(40) 69(15) H(22C) 5150(40) 7470(70) 5610(30) 42(11) H(21C) 7820(40)7310(90) 5040(40) 62(14) H(20C) 6780(30) 3790(70) 5480(30) 48(12) H(12)9030(40) 5290(90) 7280(40) 62(16)

Preparation of Amorphous Form from Form B

A Buchi Spray drier was used in this method under the followingconditions:

Inlet temperature set point: 130° C.

Outlet Temperature (start of run) 55° C.

Outlet temperature (end of run): 58° C.

Nitrogen pressure: 120 psi

Aspirator: 100%

Pump: 40%

Filter pressure 11 mbar

Condenser Temperature: 10° C.

Run Time 15 min

Yield 86.5%

Dried in 25° C. vacuum over for 24 hours.

4 g of Form B was dissolved in 86.4 g of acetone and 9.6 g water underthe above conditions. The run time was 15 min. The product was driedunder vacuum at 25° C. for over 24 hrs to produce the Amorphous Form.

The XRPD spectrum of the Amorphous Form is shown in FIG. 11.

The TGA trace for Amorphous Form is shown in FIG. 12.

The DSC trace for Amorphous Form is shown in FIG. 13.

PK and Solubility of Different Solid State Forms of Compound 1

Bioavailability of crystalline Form B, 85% amorphous Compound 1 andHPMCAS solid dispersion of Compound 1 were evaluated in rat, the resultsof which are provided in Table 4 below. These forms of the compound weredosed in an oral suspension with a vehicle containing 0.5% methylcellulose/0.5% SLS/99% water. Bioavailability of various solid form iswas evaluated as compared to a multicomponent IV solution of Compound 1.Bioavailability of crystalline polymorph B was 3-6%, compared to 61-95%for amorphous material and 109-111% for solid dispersion. In FaSSIF,crystalline polymorph B has a measured solubiity of 1.0 μg/ml, while the85% amorphous material has a solubility of 67.4 μg/ml. The crystallinematerial showed 67-74% bioavailability when dosed as a PEG solution,indicating that absorption was solubility limited.

TABLE 4 Drug Form Vehicle Dose (mg/kg) AUC (ug*hr/mL) Tmax (h) % F 85%0.5% 50 135.5 27.6 6.0 0.0 95.0 20.0 Amorphous MC/0.5% SLS 85% 0.5% 200371.9 46.1 6.0 0.0 61.0 7.0 Amorphous MC/0.5% SLS Crystalline 0.5% 508.0 1.2 4.0 0.0 5.5 0.8 MC/0.5% SLS Crystalline 0.5% 200 16.9 3.0 4.71.2 3.1 0.3 MC/0.5% SLS Crystalline PEG 50 135.1 43.0 5.5 1.0 74.0 23.0Crystalline PEG 200 431.5 101.1 14.5 11.0 67.0 16.0 Solid 0.5% 25 90.18.1 6.0 0.0 111.0 10.0 Dispersion MC/0.5% SLS Solid 0.5% 100 260.8 28.46.0 0.0 109.0 12.0 Dispersion MC/0.5% SLS

1.-77. (canceled)
 78. A pharmaceutical pack or kit comprisingsubstantially amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamideand a pharmaceutically acceptable carrier. 79.-132. (canceled)
 133. Thepharmaceutical pack or kit according to claim 78 comprisingsubstantially amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide,wherein the substantially amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidecomprises less than about 15% crystallineN-[2,4-bis(1,1-dimethylethyl)-5-hydroxypheny]-1,4-dihydro-4-oxoquinoline-3-carboxamide,and a pharmaceutically acceptable carrier.
 134. The pharmaceutical packor kit according to claim 78, wherein the substantially amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamidecomprises less than about 5% crystallineN-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide,and a pharmaceutically acceptable carrier.
 135. The pharmaceutical packor kit according to claim 78 comprising amorphousN-[2,4-bis(1,1-dimethylethyl)-5-hydroxypheny]-1,4-dihydro-4-oxoquinoline-3-carboxamideand a pharmaceutically acceptable carrier.